Pyrrolidine inhibitors of IAP

ABSTRACT

The invention provides novel inhibitors of IAP that are useful as therapeutic agents for treating malignancies where the compounds have the general formula I: 
                         
wherein A, Q, X 1 , X 2 , Y, R 1 , R 2 , R 3 , R 4 , R 4 ′, R 5 , R 6  and R 6 ′ are as described herein.

RELATED APPLICATIONS

This application is a continuation application of U.S. application Ser.No. 12/105,109, filed Apr. 17, 2008, which is a continuation applicationof U.S. application Ser. No. 11/312,063, filed Dec. 19, 2005, whichclaims priority under 35 U.S.C. §119(e)(1) to U.S. provisionalapplication 60/638,202 filed on Dec. 20, 2004 which is incorporatedherein by reference in its entirety.

FIELD OF THE INVENTION

The present invention relates to organic compounds useful for therapyand/or prophylaxis in a mammal, and in particular to inhibitors of IAPproteins useful for treating cancers.

BACKGROUND OF THE INVENTION

Apoptosis or programmed cell death is a genetically and biochemicallyregulated mechanism that plays an important role in development andhomeostasis in invertebrates as well as vertebrates. Aberrancies inapoptosis that lead to premature cell death have been linked to avariety of developmental disorders. Deficiencies in apoptosis thatresult in the lack of cell death have been linked to cancer and chronicviral infections (Thompson et al., (1995) Science 267, 1456-1462).

One of the key effector molecules in apoptosis are the caspases(cysteine containing aspartate specific proteases). Caspases are strongproteases, cleaving after aspartic acid residues and once activated,digest vital cell proteins from within the cell. Since caspases are suchstrong proteases, tight control of this family of proteins is necessaryto prevent premature cell death. In general, caspases are synthesized aslargely inactive zymogens that require proteolytic processing in orderto be active. This proteolytic processing is only one of the ways inwhich caspases are regulated. The second mechanism is through a familyof proteins that bind and inhibit caspases.

A family of molecules that inhibit caspases are the Inhibitors ofApoptosis (IAP) (Deveraux et al., J Clin Immunol (1999), 19:388-398).IAPs were originally discovered in baculovirus by their functionalability to substitute for P35 protein, an anti-apoptotic gene (Crook etal. (1993) J Virology 67, 2168-2174). IAPs have been described inorganisms ranging from Drosophila to human. Regardless of their origin,structurally, IAPs comprise one to three Baculovirus IAP repeat (BIR)domains, and most of them also possess a carboxyl-terminal RING fingermotif. The BIR domain itself is a zinc binding domain of about 70residues comprising 4 alpha-helices and 3 beta strands, with cysteineand histidine residues that coordinate the zinc ion (Hinds et al.,(1999) Nat. Struct. Biol. 6, 648-651). It is the BIR domain that isbelieved to cause the anti-apoptotic effect by inhibiting the caspasesand thus inhibiting apoptosis. As an example, human X-chromosome linkedIAP (XIAP) inhibits caspase 3, caspase 7 and the Apaf-1-cytochrome Cmediated activation of caspase 9 (Deveraux et al., (1998) EMBO J. 17,2215-2223). Caspases 3 and 7 are inhibited by the BIR2 domain of XIAP,while the BIR3 domain of XIAP is responsible for the inhibition ofcaspase 9 activity. XIAP is expressed ubiquitously in most adult andfetal tissues (Liston et al, Nature, 1996, 379(6563):349), and isoverexpressed in a number of tumor cell lines of the NCI 60 cell linepanel (Fong et al, Genomics, 2000, 70:113; Tamm et al, Clin. Cancer Res.2000, 6(5):1796). Overexpression of XIAP in tumor cells has beendemonstrated to confer protection against a variety of pro-apoptoticstimuli and promotes resistance to chemotherapy (LaCasse et al,Oncogene, 1998, 17(25):3247). Consistent with this, a strong correlationbetween XIAP protein levels and survival has been demonstrated forpatients with acute myelogenous leukemia (Tamm et al, supra).Down-regulation of XIAP expression by antisense oligonucleotides hasbeen shown to sensitize tumor cells to death induced by a wide range ofpro-apoptotic agents, both in vitro and in vivo (Sasaki et al, CancerRes., 2000, 60(20):5659; Lin et al, Biochem J., 2001, 353:299; Hu et al,Clin. Cancer Res., 2003, 9(7):2826). Smac/DIABLO-derived peptides havealso been demonstrated to sensitize a number of different tumor celllines to apoptosis induced by a variety of pro-apoptotic drugs (Arnt etal, J. Biol. Chem., 2002, 277(46):44236; Fulda et al, Nature Med., 2002,8(8):808; Guo et al, Blood, 2002, 99(9):3419; Vucic et al, J. Biol.Chem., 2002, 277(14):12275; Yang et al, Cancer Res., 2003, 63(4):831).

Melanoma IAP (ML-IAP) is an IAP not detectable in most normal adulttissues but is strongly upregulated in melanoma (Vucic et al., (2000)Current Bio 10:1359-1366).

Determination of protein structure demonstrated significant homology ofthe ML-IAP BIR and RING finger domains to corresponding domains presentin human XIAP, C-IAP1 and C-IAP2. The BIR domain of ML-IAP appears tohave the most similarities to the BIR2 and BIR3 of XIAP, C-IAP1 andC-IAP2, and appears to be responsible for the inhibition of apoptosis,as determined by deletional analysis. Furthermore, Vucic et al.,demonstrated that ML-IAP could inhibit chemotherapeutic agent inducedapoptosis. Agents such as adriamycin and 4-tertiary butylphenol (4-TBP)were tested in a cell culture system of melanomas overexpressing ML-IAPand the chemotherapeutic agents were significantly less effective inkilling the cells when compared to a normal melanocyte control. Themechanism by which ML-IAP produces an anti-apoptotic activity is in partthrough inhibition of caspase 3 and 9. ML-IAP did not effectivelyinhibit caspases 1, 2, 6, or 8.

Since apoptosis is a strictly controlled pathway with multipleinteracting factors, the discovery that IAPs themselves are regulatedwas not unusual. In the fruit fly Drosophila, the Reaper (rpr), HeadInvolution Defective (hid) and GRIM proteins physically interact withand inhibit the anti-apoptotic activity of the Drosophila family ofIAPs. In the mammal, the proteins SMAC/DIABLO act to block the IAPs andallow apoptosis to proceed. It was shown that during normal apoptosis,SMAC is processed into an active form and is released from themitochondria into the cytoplasm where it physically binds to IAPs andprevents the IAP from binding to a caspase. This inhibition of the IAPallows the caspase to remain active and thus proceed with apoptosis.Interestingly, sequence homology between the IAP inhibitors shows thatthere is a four amino acid motif in the N-terminus of the processed,active proteins. This tetrapeptide appears to bind into a hydrophobicpocket in the BIR domain and disrupts the BIR domain binding to caspases(Chai et al., (2000) Nature 406:855-862, Liu et al., (2000) Nature408:1004-1008, Wu et al., (2000) Nature 408 1008-1012).

SUMMARY OF THE INVENTION

In one aspect of the present invention there is provided novelinhibitors of IAP proteins having the general formula I:

wherein

-   -   A is a 5-member aromatic heterocycle incorporating 1 to 4        heteroartoms N, O or S and is optionally substituted with one or        more R₇ and R₈ groups;    -   Q is H, alkyl, a carbocycle, a heterocycle; wherein one or more        CH₂ or CH groups of an alkyl is optionally replaced with —O—,        —S—, —S(O)—, S(O)₂, —N(R₈)—, —C(O)—, —C(O)—NR₈—, —NR₈—C(O)—,        —SO₂—NR₈—, —NR₈—SO₂—, —NR₈—C(O)—NR₈—, —NR₈—C(NH)—NR₈—,        —NR₈—C(NH)—, —C(O)—O— or —O—C(O)—; and an alkyl, carbocycle and        heterocycle is optionally substituted with one or more hydroxyl,        alkoxy, acyl, halogen, mercapto, oxo, carboxyl, halo-substituted        alkyl, amino, cyano, nitro, amidino, guanidino an optionally        substituted carbocycle or an optionally substituted heterocycle;    -   X₁ and X₂ are each independently O or S;    -   Y is a bond, (CR₇R₇)_(n), O or S; wherein n is 1 or 2 and R₇ is        H, halogen, alkyl, aryl, aralkyl, amino, arylamino, alkylamino,        aralkylamino, alkoxy, aryloxy or aralkyloxy;    -   R₁ is H or R₁ and R₂ together form a 5-8 member heterocycle;    -   R₂ is alkyl, a carbocycle, carbocyclylalkyl, a heterocycle or        heterocyclylalkyl each optionally substituted with halogen,        hydroxyl, oxo, thione, mercapto, carboxyl, alkyl, haloalkyl,        alkoxy, alkylthio, sulfonyl, amino and nitro;    -   R₃ is H or alkyl optionally substituted with halogen or        hydroxyl; or R₃ and R₄ together form a 3-6 heterocycle;    -   R₃′ is H, or R₃ and R₃′ together form a 3-6 carbocycle;    -   R₄ and R₄′ are independently H, hydroxyl, amino, alkyl,        carbocycle, carbocycloalkyl, carbocycloalkyloxy,        carbocycloalkyloxycarbonyl, heterocycle, heterocycloalkyl,        heterocycloalkyloxy or heterocycloalkyloxycarbonyl; wherein each        alkyl, carbocycloalkyl, carbocycloalkyloxy,        carbocycloalkyloxycarbonyl, heterocycle, heterocycloalkyl,        heterocycloalkyloxy and heterocycloalkyloxycarbonyl is        optionally substituted with halogen, hydroxyl, mercapto,        carboxyl, alkyl, alkoxy, amino, imino and nitro; or R₄ and R₄′        together form a heterocycle;    -   R₅ is H or alkyl;    -   R₆, and R₆′ are each independently H, alkyl, aryl or aralkyl;    -   R₇ is H, cyano, hydroxyl, mercapto, halogen, nitro, carboxyl,        amidino, guanidino, alkyl, a carbocycle, a heterocycle or —U—V;        wherein U is —O—, —S—, —S(O)—, S(O)₂, —N(R₈)—, —C(O)—,        —C(O)—NR₈—, —NR₈—C(O)—, —SO₂—NR₈—, —NR₈—SO₂—, —NR₈—C(O)—NR₈—,        —NR₈—C(NH)—NR₈—, —NR₈—C(NH)—, —C(O)—O— or —O—C(O)— and V is        alkyl, a carbocycle or a heterocycle; and wherein one or more        CH₂ or CH groups of an alkyl is optionally replaced with —O—,        —S—, —S(O)—, S(O)₂, —N(R₈)—, —C(O)—, —C(O)—NR₈—, —NR₈—C(O)—,        —SO₂—NR₈—, —NR₈—C(O)—NR₈—, —C(O)—O— or —O—C(O)—; and an alkyl,        carbocycle and heterocycle is optionally substituted with        hydroxyl, alkoxy, acyl, halogen, mercapto, oxo, carboxyl, acyl,        halo-substituted alkyl, amino, cyano nitro, amidino, guanidino        an optionally substituted carbocycle or an optionally        substituted heterocycle;    -   R₈ is H, alkyl, a carbocycle or a heterocycle wherein one or        more CH₂ or CH groups of said alkyl is optionally replaced with        —O—, —S—, —S(O)—, S(O)₂, —N(R₈), or —C(O)—; and said alkyl,        carbocycle and heterocycle is optionally substituted with        hydroxyl, alkoxy, acyl, halogen, mercapto, oxo (═O), carboxyl,        acyl, halo-substituted alkyl, amino, cyano nitro, amidino,        guanidino an optionally substituted carbocycle or an optionally        substituted heterocycle;    -   and salts and solvates thereof.

In another aspect of the invention, there are provided compositionscomprising compounds of formula I and a carrier, diluent or excipient.

In another aspect of the invention, there is provided a method ofinducing apoptosis in a cell comprising introducing into said cell acompound of formula I.

In another aspect of the invention, there is provided a method ofsensitizing a cell to an apoptotic signal comprising introducing intosaid cell a compound of formula I.

In another aspect of the invention, there is provided a method forinhibiting the binding of an IAP protein to a caspase protein comprisingcontacting said IAP protein with a compound of formula I.

In another aspect of the invention, there is provided a method fortreating a disease or condition associated with the overexpression of anIAP protein in a mammal, comprising administering to said mammal aneffective amount of a compound of formula I.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

“Acyl” means a carbonyl containing substituent represented by theformula —C(O)—R in which R is H, alkyl, a carbocycle, a heterocycle,carbocycle-substituted alkyl or heterocycle-substituted alkyl whereinthe alkyl, alkoxy, carbocycle and heterocycle are as defined herein.Acyl groups include alkanoyl (e.g. acetyl), aroyl (e.g. benzoyl), andheteroaroyl.

“Alkyl” means a branched or unbranched, saturated or unsaturated (i.e.alkenyl, alkynyl) aliphatic hydrocarbon group, having up to 12 carbonatoms unless otherwise specified. When used as part of another term, forexample “alkylamino”, the alkyl portion may be a saturated hydrocarbonchain, however also includes unsaturated hydrocarbon carbon chains suchas “alkenylamino” and “alkynylamino. Examples of particular alkyl groupsare methyl, ethyl, n-propyl, isopropyl, n-butyl, iso-butyl, sec-butyl,tert-butyl, n-pentyl, 2-methylbutyl, 2,2-dimethylpropyl, n-hexyl,2-methylpentyl, 2,2-dimethylbutyl, n-heptyl, 3-heptyl, 2-methylhexyl,and the like. The terms “lower alkyl” “C₁-C₄ alkyl” and “alkyl of 1 to 4carbon atoms” are synonymous and used interchangeably to mean methyl,ethyl, 1-propyl, isopropyl, cyclopropyl, 1-butyl, sec-butyl or t-butyl.Unless specified, substituted, alkyl groups may contain one, for exampletwo, three or four substituents which may be the same or different.Examples of substituents are, unless otherwise defined, halogen, amino,hydroxyl, protected hydroxyl, mercapto, carboxy, alkoxy, nitro, cyano,amidino, guanidino, urea, sulfonyl, sulfinyl, aminosulfonyl,alkylsulfonylamino, arylsulfonylamino, aminocarbonyl, acylamino, alkoxy,acyl, acyloxy, a carbocycle, a heterocycle. Examples of the abovesubstituted alkyl groups include, but are not limited to; cyanomethyl,nitromethyl, hydroxymethyl, trityloxymethyl, propionyloxymethyl,aminomethyl, carboxymethyl, carboxyethyl, carboxypropyl,alkyloxycarbonylmethyl, allyloxycarbonylaminomethyl, carbamoyloxymethyl,methoxymethyl, ethoxymethyl, t-butoxymethyl, acetoxymethyl,chloromethyl, bromomethyl, iodomethyl, trifluoromethyl, 6-hydroxyhexyl,2,4-dichloro(n-butyl), 2-amino(iso-propyl), 2-carbamoyloxyethyl and thelike. The alkyl group may also be substituted with a carbocycle group.Examples include cyclopropylmethyl, cyclobutylmethyl, cyclopentylmethyl,and cyclohexylmethyl groups, as well as the corresponding -ethyl,-propyl, -butyl, -pentyl, -hexyl groups, etc. Substituted alkyls includesubstituted methyls e.g. a methyl group substituted by the samesubstituents as the “substituted C_(n)-C_(m) alkyl” group. Examples ofthe substituted methyl group include groups such as hydroxymethyl,protected hydroxymethyl (e.g. tetrahydropyranyloxymethyl),acetoxymethyl, carbamoyloxymethyl, trifluoromethyl, chloromethyl,carboxymethyl, bromomethyl and iodomethyl.

“Amidine” means the group —C(NH)—NHR wherein R is H or alkyl or aralkyl.A particular amidine is the group —NH—C(NH)—NH₂.

“Amino” means primary (i.e. —NH₂), secondary (i.e. —NRH) and tertiary(i.e. —NRR) amines. Particular secondary and tertiary amines arealkylamine, dialkylamine, arylamine, diarylamine, aralkylamine anddiaralkylamine wherein the alkyl is as herein defined and optionallysubstituted. Particular secondary and tertiary amines are methylamine,ethylamine, propylamine, isopropylamine, phenylamine, benzylaminedimethylamine, diethylamine, dipropylamine and disopropylamine.

“Amino-protecting group” as used herein refers to a derivative of thegroups commonly employed to block or protect an amino group whilereactions are carried out on other functional groups on the compound.Examples of such protecting groups include carbamates, amides, alkyl andaryl groups, imines, as well as many N-heteroatom derivatives which canbe removed to regenerate the desired amine group. Particular aminoprotecting groups are Boc, Fmoc and Cbz. Further examples of thesegroups are found in T. W. Greene and P. G. M. Wuts, “Protective Groupsin Organic Synthesis”, 2^(nd) ed., John Wiley & Sons, Inc., New York,N.Y., 1991, chapter 7; E. Haslam, “Protective Groups in OrganicChemistry”, J. G. W. McOmie, Ed., Plenum Press, New York, N.Y., 1973,Chapter 5, and T. W. Greene, “Protective Groups in Organic Synthesis”,John Wiley and Sons, New York, N.Y., 1981. The term “protected amino”refers to an amino group substituted with one of the aboveamino-protecting groups.

“Aryl” when used alone or as part of another term means a carbocyclicaromatic group whether or not fused having the number of carbon atomsdesignated or if no number is designated, up to 14 carbon atoms.Particular aryl groups are phenyl, naphthyl, biphenyl, phenanthrenyl,naphthacenyl, and the like (see e.g. Lang's Handbook of Chemistry (Dean,J. A., ed) 13^(th) ed. Table 7-2 [1985]). A particular aryl is phenyl.Substituted phenyl or substituted aryl means a phenyl group or arylgroup substituted with one, two, three, four or five, for example 1-2,1-3 or 1-4 substituents chosen, unless otherwise specified, from halogen(F, Cl, Br, I), hydroxy, protected hydroxy, cyano, nitro, alkyl (forexample C₁-C₆ alkyl), alkoxy (for example C₁-C₆ alkoxy), benzyloxy,carboxy, protected carboxy, carboxymethyl, protected carboxymethyl,hydroxymethyl, protected hydroxymethyl, aminomethyl, protectedaminomethyl, trifluoromethyl, alkylsulfonylamino,alkylsulfonylaminoalkyl, arylsulfonylamino, arylsulonylaminoalkyl,heterocyclylsulfonylamino, heterocyclylsulfonylaminoalkyl, heterocyclyl,aryl, or other groups specified. One or more methyne (CH) and/ormethylene (CH₂) groups in these substituents may in turn be substitutedwith a similar group as those denoted above. Examples of the term“substituted phenyl” includes but is not limited to a mono- ordi(halo)phenyl group such as 2-chlorophenyl, 2-bromophenyl,4-chlorophenyl, 2,6-dichlorophenyl, 2,5-dichlorophenyl,3,4-dichlorophenyl, 3-chlorophenyl, 3-bromophenyl, 4-bromophenyl,3,4-dibromophenyl, 3-chloro-4-fluorophenyl, 2-fluorophenyl and the like;a mono- or di(hydroxy)phenyl group such as 4-hydroxyphenyl,3-hydroxyphenyl, 2,4-dihydroxyphenyl, the protected-hydroxy derivativesthereof and the like; a nitrophenyl group such as 3- or 4-nitrophenyl; acyanophenyl group, for example, 4-cyanophenyl; a mono- or di(loweralkyl)phenyl group such as 4-methylphenyl, 2,4-dimethylphenyl,2-methylphenyl, 4-(iso-propyl)phenyl, 4-ethylphenyl, 3-(n-propyl)phenyland the like; a mono or di(alkoxy)phenyl group, for example,3,4-dimethoxyphenyl, 3-methoxy-4-benzyloxyphenyl,3-methoxy-4-(1-chloromethyl)benzyloxy-phenyl, 3-ethoxyphenyl,4-(isopropoxy)phenyl, 4-(t-butoxy)phenyl, 3-ethoxy-4-methoxyphenyl andthe like; 3- or 4-trifluoromethylphenyl; a mono- or dicarboxyphenyl or(protected carboxy)phenyl group such 4-carboxyphenyl; a mono- ordi(hydroxymethyl)phenyl or (protected hydroxymethyl)phenyl such as3-(protected hydroxymethyl)phenyl or 3,4-di(hydroxymethyl)phenyl; amono- or di(aminomethyl)phenyl or (protected aminomethyl)phenyl such as2-(aminomethyl)phenyl or 2,4-(protected aminomethyl)phenyl; or a mono-or di(N-(methylsulfonylamino))phenyl such as3-(N-methylsulfonylamino))phenyl. Also, the term “substituted phenyl”represents disubstituted phenyl groups where the substituents aredifferent, for example, 3-methyl-4-hydroxyphenyl,3-chloro-4-hydroxyphenyl, 2-methoxy-4-bromophenyl,4-ethyl-2-hydroxyphenyl, 3-hydroxy-4-nitrophenyl,2-hydroxy-4-chlorophenyl, and the like, as well as trisubstituted phenylgroups where the substituents are different, for example3-methoxy-4-benzyloxy-6-methyl sulfonylamino,3-methoxy-4-benzyloxy-6-phenyl sulfonylamino, and tetrasubstitutedphenyl groups where the substituents are different such as3-methoxy-4-benzyloxy-5-methyl-6-phenyl sulfonylamino. Particularsubstituted phenyl groups include the 2-chlorophenyl, 2-aminophenyl,2-bromophenyl, 3-methoxyphenyl, 3-ethoxy-phenyl, 4-benzyloxyphenyl,4-methoxyphenyl, 3-ethoxy-4-benzyloxyphenyl, 3,4-diethoxyphenyl,3-methoxy-4-benzyloxyphenyl,3-methoxy-4-(1-chloromethyl)benzyloxy-phenyl,3-methoxy-4-(1-chloromethyl)benzyloxy -6-methyl sulfonyl aminophenylgroups. Fused aryl rings may also be substituted with any, for example1, 2 or 3, of the substituents specified herein in the same manner assubstituted alkyl groups.

“Carbocyclyl”, “carbocyclylic”, “carbocycle” and “carbocyclo” alone andwhen used as a moiety in a complex group such as a carbocycloalkylgroup, refers to a mono-, bi-, or tricyclic aliphatic ring having 3 to14 carbon atoms, for example 3 to 7 carbon atoms, which may be saturatedor unsaturated, aromatic or non-aromatic. Particular saturatedcarbocyclic groups are cyclopropyl, cyclobutyl, cyclopentyl andcyclohexyl groups. A particular saturated carbocycle is cyclopropyl.Another particular saturated carbocycle is cyclohexyl. Particularunsaturated carbocycles are aromatic e.g. aryl groups as previouslydefined, for example phenyl. The terms “substituted carbocyclyl”,“carbocycle” and “carbocyclo” mean these groups substituted by the samesubstituents as the “substituted alkyl” group.

“Carboxy-protecting group” as used herein refers to one of the esterderivatives of the carboxylic acid group commonly employed to block orprotect the carboxylic acid group while reactions are carried out onother functional groups on the compound. Examples of such carboxylicacid protecting groups include 4-nitrobenzyl, 4-methoxybenzyl,3,4-dimethoxybenzyl, 2,4-dimethoxybenzyl, 2,4,6-trimethoxybenzyl,2,4,6-trimethylbenzyl, pentamethylbenzyl, 3,4-methylenedioxybenzyl,benzhydryl, 4,4′-dimethoxybenzhydryl, 2,2′,4,4′-tetramethoxybenzhydryl,alkyl such as t-butyl or t-amyl, trityl, 4-methoxytrityl,4,4′-dimethoxytrityl, 4,4′,4″-trimethoxytrityl, 2-phenylprop-2-yl,trimethylsilyl, t-butyldimethylsilyl, phenacyl, 2,2,2-trichloroethyl,beta-(trimethylsilyl)ethyl, beta-(di(n-butyl)methylsilyl)ethyl,p-toluenesulfonylethyl, 4-nitrobenzylsulfonylethyl, allyl, cinnamyl,1-(trimethylsilylmethyl)prop-1-en-3-yl, and like moieties. The speciesof carboxy-protecting group employed is not critical so long as thederivatized carboxylic acid is stable to the condition of subsequentreaction(s) on other positions of the molecule and can be removed at theappropriate point without disrupting the remainder of the molecule. Inparticular, it is important not to subject a carboxy-protected moleculeto strong nucleophilic bases, such as lithium hydroxide or NaOH, orreductive conditions employing highly activated metal hydrides such asLiAlH₄. (Such harsh removal conditions are also to be avoided whenremoving amino-protecting groups and hydroxy-protecting groups,discussed below.) Particular carboxylic acid protecting groups are thealkyl (e.g. methyl, ethyl, t-butyl), allyl, benzyl and p-nitrobenzylgroups. Similar carboxy-protecting groups used in the cephalosporin,penicillin and peptide arts can also be used to protect a carboxy groupsubstituents. Further examples of these groups are found in T. W. Greeneand P. G. M. Wuts, “Protective Groups in Organic Synthesis”, 2^(nd) ed.,John Wiley & Sons, Inc., New York, N.Y., 1991, chapter 5; E. Haslam,“Protective Groups in Organic Chemistry”, J. G. W. McOmie, Ed., PlenumPress, New York, N.Y., 1973, Chapter 5, and T. W. Greene, “ProtectiveGroups in Organic Synthesis”, John Wiley and Sons, New York, N.Y., 1981,Chapter 5. The term “protected carboxy” refers to a carboxy groupsubstituted with one of the above carboxy-protecting groups.

“Guanidine” means the group —NH—C(NH)—NHR wherein R is H or alkyl oraralkyl. A particular guanidine is the group —NH—C(NH)—NH₂.

“Hydroxy-protecting group” as used herein refers to a derivative of thehydroxy group commonly employed to block or protect the hydroxy groupwhile reactions are carried out on other functional groups on thecompound. Examples of such protecting groups includetetrahydropyranyloxy, benzoyl, acetoxy, carbamoyloxy, benzyl, andsilylethers (e.g. TBS, TBDPS) groups. Further examples of these groupsare found in T. W. Greene and P. G. M. Wuts, “Protective Groups inOrganic Synthesis”, 2^(nd) ed., John Wiley & Sons, Inc., New York, N.Y.,1991, chapters 2-3; E. Haslam, “Protective Groups in Organic Chemistry”,J. G. W. McOmie, Ed., Plenum Press, New York, N.Y., 1973, Chapter 5, andT. W. Greene, “Protective Groups in Organic Synthesis”, John Wiley andSons, New York, N.Y., 1981. The term “protected hydroxy” refers to ahydroxy group substituted with one of the above hydroxy-protectinggroups.

“Heterocyclic group”, “heterocyclic”, “heterocycle”, “heterocyclyl”, or“heterocyclo” alone and when used as a moiety in a complex group such asa heterocycloalkyl group, are used interchangeably and refer to anymono-, bi-, or tricyclic, saturated or unsaturated, aromatic(heteroaryl) or non-aromatic ring having the number of atoms designated,generally from 5 to about 14 ring atoms, where the ring atoms are carbonand at least one heteroatom (nitrogen, sulfur or oxygen), for example 1to 4 heteroatoms. Typically, a 5-membered ring has 0 to 2 double bondsand 6- or 7-membered ring has 0 to 3 double bonds and the nitrogen orsulfur heteroatoms may optionally be oxidized (e.g. SO, SO₂), and anynitrogen heteroatom may optionally be quaternized. Particularnon-aromatic heterocycles are morpholinyl (morpholino), pyrrolidinyl,oxiranyl, oxetanyl, tetrahydrofuranyl, 2,3-dihydrofuranyl, 2H-pyranyl,tetrahydropyranyl, thiiranyl, thietanyl, tetrahydrothietanyl,aziridinyl, azetidinyl, 1-methyl-2-pyrrolyl, piperazinyl andpiperidinyl. A “heterocycloalkyl” group is a heterocycle group asdefined above covalently bonded to an alkyl group as defined above.Particular 5-membered heterocycles containing a sulfur or oxygen atomand one to three nitrogen atoms are thiazolyl, in particularthiazol-2-yl and thiazol-2-yl N-oxide, thiadiazolyl, in particular1,3,4-thiadiazol-5-yl and 1,2,4-thiadiazol-5-yl, oxazolyl, for exampleoxazol-2-yl, and oxadiazolyl, such as 1,3,4-oxadiazol-5-yl, and1,2,4-oxadiazol-5-yl. Particular 5-membered ring heterocycles containing2 to 4 nitrogen atoms include imidazolyl, such as imidazol-2-yl;triazolyl, such as 1,3,4-triazol-5-yl; 1,2,3-triazol-5-yl,1,2,4-triazol-5-yl, and tetrazolyl, such as 1H-tetrazol-5-yl. Particularbenzo-fused 5-membered heterocycles are benzoxazol-2-yl,benzthiazol-2-yl and benzimidazol-2-yl. Particular 6-memberedheterocycles contain one to three nitrogen atoms and optionally a sulfuror oxygen atom, for example pyridyl, such as pyrid-2-yl, pyrid-3-yl, andpyrid-4-yl; pyrimidyl, such as pyrimid-2-yl and pyrimid-4-yl; triazinyl,such as 1,3,4-triazin-2-yl and 1,3,5-triazin-4-yl; pyridazinyl, inparticular pyridazin-3-yl, and pyrazinyl. The pyridine N-oxides andpyridazine N-oxides and the pyridyl, pyrimid-2-yl, pyrimid-4-yl,pyridazinyl and the 1,3,4-triazin-2-yl groups, are a particular group.Substituents for “optionally substituted heterocycles”, and furtherexamples of the 5- and 6-membered ring systems discussed above can befound in W. Druckheimer et al, U.S. Pat. No. 4,278,793. In a particularembodiment, such optionally substituted heterocycle groups aresubstituted with hydroxyl, alkyl, alkoxy, acyl, halogen, mercapto, oxo,carboxyl, acyl, halo-substituted alkyl, amino, cyano, nitro, amidino andguanidino.

“Heteroaryl” alone and when used as a moiety in a complex group such asa heteroaralkyl group, refers to any mono-, bi-, or tricyclic aromaticring system having the number of atoms designated where at least onering is a 5-, 6- or 7-membered ring containing from one to fourheteroatoms selected from the group nitrogen, oxygen, and sulfur, and ina particular embodiment at least one heteroatom is nitrogen (Lang'sHandbook of Chemistry, supra). Included in the definition are anybicyclic groups where any of the above heteroaryl rings are fused to abenzene ring. Particular heteroaryls incorporate a nitrogen or oxygenheteroatom. The following ring systems are examples of the heteroaryl(whether substituted or unsubstituted) groups denoted by the term“heteroaryl”: thienyl, furyl, imidazolyl, pyrazolyl, thiazolyl,isothiazolyl, oxazolyl, isoxazolyl, triazolyl, thiadiazolyl,oxadiazolyl, tetrazolyl, thiatriazolyl, oxatriazolyl, pyridyl,pyrimidyl, pyrazinyl, pyridazinyl, thiazinyl, oxazinyl, triazinyl,thiadiazinyl, oxadiazinyl, dithiazinyl, dioxazinyl, oxathiazinyl,tetrazinyl, thiatriazinyl, oxatriazinyl, dithiadiazinyl, imidazolinyl,dihydropyrimidyl, tetrahydropyrimidyl, tetrazolo[1,5-b]pyridazinyl andpurinyl, as well as benzo-fused derivatives, for example benzoxazolyl,benzofuryl, benzothiazolyl, benzothiadiazolyl, benzotriazolyl,benzoimidazolyl and indolyl. A particular “heteroaryl” is:1,3-thiazol-2-yl, 4-(carboxymethyl)-5-methyl-1,3-thiazol-2-yl,4-(carboxymethyl)-5-methyl-1,3-thiazol-2-yl sodium salt,1,2,4-thiadiazol-5-yl, 3-methyl-1,2,4-thiadiazol-5-yl,1,3,4-triazol-5-yl, 2-methyl-1,3,4-triazol-5-yl,2-hydroxy-1,3,4-triazol-5-yl, 2-carboxy-4-methyl-1,3,4-triazol-5-ylsodium salt, 2-carboxy-4-methyl-1,3,4-triazol-5-yl, 1,3-oxazol-2-yl,1,3,4-oxadiazol-5-yl, 2-methyl-1,3,4-oxadiazol-5-yl,2-(hydroxymethyl)-1,3,4-oxadiazol-5-yl, 1,2,4-oxadiazol-5-yl,1,3,4-thiadiazol-5-yl, 2-thiol-1,3,4-thiadiazol-5-yl,2-(methylthio)-1,3,4-thiadiazol-5-yl, 2-amino-1,3,4-thiadiazol-5-yl,1H-tetrazol-5-yl, 1-methyl-1H-tetrazol-5-yl,1-(1-(dimethylamino)eth-2-yl)-1H-tetrazol-5-yl,1-(carboxymethyl)-1H-tetrazol-5-yl, 1-(carboxymethyl)-1H-tetrazol-5-ylsodium salt, 1-(methylsulfonic acid)-1H-tetrazol-5-yl, 1-(methylsulfonicacid)-1H-tetrazol-5-yl sodium salt, 2-methyl-1H-tetrazol-5-yl,1,2,3-triazol-5-yl, 1-methyl-1,2,3-triazol-5-yl,2-methyl-1,2,3-triazol-5-yl, 4-methyl-1,2,3-triazol-5-yl, pyrid-2-ylN-oxide, 6-methoxy-2-(n-oxide)-pyridaz-3-yl, 6-hydroxypyridaz-3-yl,1-methylpyrid-2-yl, 1-methylpyrid-4-yl, 2-hydroxypyrimid-4-yl,1,4,5,6-tetrahydro-5,6-dioxo-4-methyl-as-triazin-3-yl,1,4,5,6-tetrahydro-4-(formylmethyl)-5,6-dioxo-as-triazin-3-yl,2,5-dihydro-5-oxo-6-hydroxy-astriazin-3-yl,2,5-dihydro-5-oxo-6-hydroxy-as-triazin-3-yl sodium salt,2,5-dihydro-5-oxo-6-hydroxy-2-methyl-astriazin-3-yl sodium salt,2,5-dihydro-5-oxo-6-hydroxy-2-methyl-as-triazin-3-yl,2,5-dihydro-5-oxo-6-methoxy-2-methyl-as-triazin-3-yl,2,5-dihydro-5-oxo-as-triazin-3-yl,2,5-dihydro-5-oxo-2-methyl-as-triazin-3-yl,2,5-dihydro-5-oxo-2,6-dimethyl-as-triazin-3-yl,tetrazolo[1,5-b]pyridazin-6-yl and8-aminotetrazolo[1,5-b]-pyridazin-6-yl. An alternative group of“heteroaryl” includes; 4-(carboxymethyl)-5-methyl-1,3-thiazol-2-yl,4-(carboxymethyl)-5-methyl-1,3-thiazol-2-yl sodium salt,1,3,4-triazol-5-yl, 2-methyl-1,3,4-triazol-5-yl, 1H-tetrazol-5-yl,1-methyl-1H-tetrazol-5-yl,1-(1-(dimethylamino)eth-2-yl)-1H-tetrazol-5-yl,1-(carboxymethyl)-1H-tetrazol-5-yl, 1-(carboxymethyl)-1H-tetrazol-5-ylsodium salt, 1-(methylsulfonic acid)-1H-tetrazol-5-yl, 1-(methylsulfonicacid)-1H-tetrazol-5-yl sodium salt, 1,2,3-triazol-5-yl,1,4,5,6-tetrahydro-5,6-dioxo-4-methyl-as-triazin-3-yl,1,4,5,6-tetrahydro-4-(2-formylmethyl)-5,6-dioxo-as-triazin-3-yl,2,5-dihydro-5-oxo-6-hydroxy-2-methyl-as-triazin-3-yl sodium salt,2,5-dihydro-5-oxo-6-hydroxy-2-methyl-as-triazin-3-yl,tetrazolo[1,5-b]pyridazin-6-yl, and8-aminotetrazolo[1,5-b]pyridazin-6-yl. Heteroaryl groups are optionallysubstituted as described for heterocycles.

“Inhibitor” means a compound which reduces or prevents the binding ofIAP proteins to caspase proteins or which reduces or prevents theinhibition of apoptosis by an IAP protein. Alternatively, “inhibitor”means a compound which prevents the binding interaction of X-IAP withcaspases or the binding interaction of ML-IAP with SMAC.

“Optionally substituted” unless otherwise specified means that a groupmay be substituted by one or more (e.g. 0, 1, 2, 3 or 4) of thesubstituents listed for that group in which said substituents may be thesame or different. In an embodiment an optionally substituted group has1 substituent. In another embodiment an optionally substituted group has2 substituents. In another embodiment an optionally substituted grouphas 3 substituents.

“Pharmaceutically acceptable salts” include both acid and base additionsalts. “Pharmaceutically acceptable acid addition salt” refers to thosesalts which retain the biological effectiveness and properties of thefree bases and which are not biologically or otherwise undesirable,formed with inorganic acids such as hydrochloric acid, hydrobromic acid,sulfuric acid, nitric acid, carbonic acid, phosphoric acid and the like,and organic acids may be selected from aliphatic, cycloaliphatic,aromatic, araliphatic, heterocyclic, carboxylic, and sulfonic classes oforganic acids such as formic acid, acetic acid, propionic acid, glycolicacid, gluconic acid, lactic acid, pyruvic acid, oxalic acid, malic acid,maleic acid, maloneic acid, succinic acid, fumaric acid, tartaric acid,citric acid, aspartic acid, ascorbic acid, glutamic acid, anthranilicacid, benzoic acid, cinnamic acid, mandelic acid, embonic acid,phenylacetic acid, methanesulfonic acid, ethanesulfonic acid,p-toluenesulfonic acid, salicyclic acid and the like.

“Pharmaceutically acceptable base addition salts” include those derivedfrom inorganic bases such as sodium, potassium, lithium, ammonium,calcium, magnesium, iron, zinc, copper, manganese, aluminum salts andthe like. Particularly base addition salts are the ammonium, potassium,sodium, calcium and magnesium salts. Salts derived from pharmaceuticallyacceptable organic nontoxic bases includes salts of primary, secondary,and tertiary amines, substituted amines including naturally occurringsubstituted amines, cyclic amines and basic ion exchange resins, such asisopropylamine, trimethylamine, diethylamine, triethylamine,tripropylamine, ethanolamine, 2-diethylaminoethanol, trimethamine,dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine,hydrabamine, choline, betaine, ethylenediamine, glucosamine,methylglucamine, theobromine, purines, piperizine, piperidine,N-ethylpiperidine, polyamine resins and the like. Particularly organicnon-toxic bases are isopropylamine, diethylamine, ethanolamine,trimethamine, dicyclohexylamine, choline, and caffeine.

“Sulfonyl” means a —SO₂—R group wherein R is alkyl, carbocycle,heterocycle, carbocycloalkyl or heterocycloalkyl. Particular sulfonylgroups are alkylsulfonyl (i.e. —SO₂— alkyl), for example methylsulfonyl;arylsulfonyl, for example phenylsulfonyl; aralkylsulfonyl, for examplebenzylsulfonyl.

The phrase “and salts and solvates thereof” as used herein means thatcompounds of the inventions may exist in one or a mixture of salts andsolvate forms. For example a compound of the invention may besubstantially pure in one particular salt or solvate form or else may bemixtures of two or more salt or solvate forms.

The present invention provides novel compounds having the generalformula I:

wherein A, Q, X₁, X₂, Y, R₁, R₂, R₃, R₄, R₄′, R₅, R₆ and R₆′ are asdescribed herein.

Ring A is a 5-member aromatic heterocycle incorporating 1 to 4heteroartoms N, O or S which is substituted with group Q and isoptionally further substituted with one or more R₇ (for substitutions ata ring carbon atom) and one or more R₈ (for substitutions at a ringnitrogen).

R₇ in each occurrence is independently H, cyano, hydroxyl, mercapto,halogen, nitro, carboxyl, amidino, guanidino, alkyl, a carbocycle, aheterocycle or —U—V; wherein U is —O—, —S—, —S(O)—, S(O)₂, —N(R₈)—,—C(O)—, —C(O)—NR₈—, —NR₈—C(O)—, —SO₂—NR₈—, —NR₈—SO₂—, —NR₈—C(O)—NR₈—,—NR₈—C(NH)—NR₈—, —NR₈—C(NH)—, —C(O)—O— or —O—C(O)— and V is alkyl, acarbocycle or a heterocycle; and wherein one or more CH₂ or CH groups ofan alkyl is optionally replaced with —O—, —S—, —S(O)—, S(O)₂, —N(R₈)—,—C(O)—, —C(O)—NR₈—, —NR₈—C(O)—, —SO₂—NR₈—, —NR₈—SO₂—, —NR₈—C(O)—NR₈—,—NR₈—C(NH)—NR₈—, —NR₈—C(NH)—, —C(O)—O— or —O—C(O)—; and an alkyl,carbocycle and heterocycle is optionally substituted with hydroxyl,alkoxy, acyl, halogen, mercapto, oxo, carboxyl, acyl, halo-substitutedalkyl, amino, cyano, nitro, amidino, guanidino an optionally substitutedcarbocycle or an optionally substituted heterocycle. Substituents of the“optionally substituted carbocycle” and “optionally substitutedheterocycle” are as defined herein. In a particular embodiment suchcarbocycle and heterocycle groups are substituted with hydroxyl, alkyl,alkoxy, acyl, halogen, mercapto, oxo, carboxyl, acyl, halo-substitutedalkyl, amino, cyano, nitro, amidino and guanidino. In an embodiment R₇is H, halogen, cyano, alkyl, hydroxyalkyl or alkoxyalkyl.

R₈ is H, alkyl, a carbocycle or a heterocycle wherein one or more CH₂ orCH groups of said alkyl is optionally replaced with —O—, —S—, —S(O)—,S(O)₂, —N(R₈), or —C(O)—; and said alkyl, carbocycle and heterocycle isoptionally substituted with hydroxyl, alkoxy, acyl, halogen, mercapto,oxo (═O), carboxyl, acyl, halo-substituted alkyl, amino, cyano nitro,amidino, guanidino an optionally substituted carbocycle or an optionallysubstituted heterocycle. Substituents of the “optionally substitutedcarbocycle” and “optionally substituted heterocycle” are as definedherein. In a particular embodiment such carbocycle and heterocyclegroups are substituted with hydroxyl, alkyl, alkoxy, acyl, halogen,mercapto, oxo, carboxyl, acyl, halo-substituted alkyl, amino, cyano,nitro, amidino and guanidino. In a particular embodiment R₈ is H, alkyl,or acyl. In an embodiment R₈ is methyl. In another embodiment R₈ isacetyl. In a particular embodiment R₈ is H. In an embodiment R₇ is H,halogen, amino, hydroxyl, carboxyl, alkyl, haloalkyl or aralkyl. In aparticular embodiment R₇ is halogen, for example Cl or F. In aparticular embodiment R₇ is H. It is understood that substitutionsdefined for R₇ and R₈ as well as all other variable groups herein aresubject to permissible valency.

In a particular embodiment ring A has the general formula II:

wherein Z₁ is NR₈, O or S; and Z₂, Z₃ and Z₄ are each independently N orCR₇. Group Q is attached to ring A of formula II and II′ at the ringmember between Z₂ and Z₃. In a particular embodiment Z₁ is S. In aparticular embodiment Z₁ is O. In another particular embodiment Z₁ isNR₈ wherein R₈ is as defined herein. In a particular embodiment Z₁ isNR₈ wherein R₈ is H. In another particular embodiment Z₁ is NR₈ whereinR₈ is Me. In another embodiment Z₁ is O or S while Z₂ is N and Z₃ is Nor CR₇. In a particular embodiment Z₁ is S while Z₂ is N and Z₃ is CR₇.In a particular embodiment Z₁ is S while Z₂ is N and Z₃ is CH.

In a particular embodiment, ring A is an aromatic heterocyle selectedfrom the group consisting of IIa-IIcc:

wherein R₇ and R₈ are as defined herein. Q is not part of ring A and isshown for positional purposes. In a particular embodiment, ring A is anyone of the groups IIa-IIz wherein R₈ is H and R₇ is H, Cl, orhydroxypropynyl. In another particular embodiment, ring A is any one ofthe groups IIa-IIz wherein R₇ and R₈ are both H. In another embodiment,ring A is the IIg. In another embodiment, ring A is IIg and R₇ is H.

Q is H, alkyl, a carbocycle, a heterocycle; wherein one or more CH₂ orCH groups of an alkyl is optionally replaced with —O—, —S—, —S(O)—,S(O)₂, —N(R₈)—, —C(O)—, —C(O)—NR₈—, —NR₈—C(O)—, —SO₂—NR₈—, —NR₈—SO₂—,—NR₈—C(O)—NR₈—, —NR₈—C(NH)—NR₈—, —NR₈—C(NH)—, —C(O)—O— or —O—C(O)—; andwherein any of the foregoing alkyl, carbocycle and heterocycle isoptionally substituted with one or more hydroxyl, alkoxy, acyl, halogen,mercapto, oxo, carboxyl, acyl, halo-substituted alkyl, amino, cyanonitro, amidino, guanidino an optionally substituted carbocycle or anoptionally substituted heterocycle. Substituents of the “optionallysubstituted carbocycle” and “optionally substituted heterocycle” are asdefined herein. In a particular embodiment such carbocycle andheterocycle groups are substituted with hydroxyl, alkyl, alkoxy, acyl,halogen, mercapto, oxo, carboxyl, acyl, halo-substituted alkyl, amino,cyano, nitro, amidino and guanidino. In a particular embodiment Q is acarbocycle or heterocycle optionally substituted with halogen, amino,oxo, alkyl, a carbocycle or a heterocycle; wherein one or more CH₂ or CHgroups of an alkyl is optionally replaced with —O—, —S—, —S(O)—, S(O)₂,—N(R₈)—, —C(O)—, —C(O)—NR₈—, —NR₈—C(O)—, —SO₂NR₈—, —NR₈—SO₂—,—NR₈—C(O)—NR₈—, —NR₈—C(NH)—NR₈—, —NR₈—C(NH)—, —C(O)—O— or —O—C(O)—; andwherein said alkyl, carbocycle or heterocycle is optionally substitutedwith halogen, amino, hydroxyl, mercapto, carboxyl, alkoxy, alkoxyalkoxy,hydroxyalkoxy, alkylthio, acyloxy, acyloxyalkoxy, alkylsulfonyl,alkylsulfonylalkyl, alkylsulfinyl, and alkylsulfinylalkyl.

In a particular embodiment, Q is a carbocycle or heterocycle selectedfrom the group consisting of IIIa-IIIs:

wherein n is 1-4, for example 1-3, for example 1-2, for example 1; T isO, S, NR₈ or CR₇R₇; W is O, NR₈ or CR₇R₇; and R₇ and R₈ are as definedherein. In a particular embodiment Q is any one of IIIa-IIIi wherein R₈is H and R₇ is selected from the group consisting of H, F, Cl, Me,methoxy, hydroxyethoxy, methoxyethoxy, acetoxyethoxy, methylsulfonylmethylsulfonylmethyl, phenyl and morpholin-4-yl. In another particularembodiment Q is IIId. In a particular embodiment Q is IIId which issubstituted at the 4-position with R₇. In another particular embodimentQ is IIId which is substituted at the 5-position with R₇.

X₁ and X₂ are each independently O or S. In a particular embodiment, X₁and X₂ are both O. In another particular embodiment X₁ and X₂ are bothS. In another particular embodiment, X₁ is S while X₂ is O. In anotherparticular embodiment, X₁ is O while X₂ is S.

Y is a bond, (CR₇R₇)_(n), O or S; wherein n is 1 or 2 and R₇ is H,halogen, alkyl, aryl, aralkyl, amino, arylamino, alkylamino,aralkylamino, alkoxy, aryloxy or aralkyloxy. In a particular embodiment,Y is (CHR₇)_(n), O or S; wherein n is 1 or 2 and R₇ is H, halogen,alkyl, aryl, aralkyl, amino, arylamino, alkylamino, aralkylamino,alkoxy, aryloxy or aralkyloxy. In a particular embodiment, Y is CH₂. Ina particular embodiment n is 1. In a particular embodiment Y is a bond.In a particular embodiment n is 1 and Y is CHR₇ wherein R₇ isaralkyloxy, for example benzyloxy. In a particular embodiment n is 1 andY is CHR₇ wherein R₇ is F. In a particular embodiment n is 1 and Y isCHR₇ wherein R₇ is aralkylamino, for example benzylamino. In anotherparticular embodiment Y is O. In another particular embodiment Y is S.

R₁ is H or R₁ and R₂ together form a 5-8 member ring. In a particularembodiment, R₁ is H. In a particular embodiment, R₁ and R₂ together forma 6-member ring. In a particular embodiment, R₁ and R₂ together form a7-member ring. In another particular embodiment, R₁ and R₂ together forman 8-member ring. In another particular embodiment, R₁ and R₂ togetherform a 7-member ring while Y is S. In another particular embodiment, R₁is H, while Y is CH₂. In another particular embodiment, R₁ is H, while Yis S. In another particular embodiment, R₁ is H, while Y is O.

R₂ is alkyl, a carbocycle, carbocyclylalkyl, a heterocycle orheterocyclylalkyl each optionally substituted with halogen, hydroxyl,oxo, thione, mercapto, carboxyl, alkyl, haloalkyl, alkoxy, alkylthio,sulfonyl, amino and nitro. In a particular embodiment R₂ is alkyl, acarbocycle, carbocyclylalkyl, a heterocycle or heterocyclylalkyl eachoptionally substituted with halogen, hydroxyl, oxo, mercapto, thione,carboxyl, alkyl, haloalkyl, alkoxy, alkylthio, sulfonyl, amino andnitro. In an embodiment R₂ is alkyl, a carbocycle, carbocyclylalkyl, aheterocycle or heterocyclylalkyl each optionally substituted withhalogen, hydroxyl, mercapto, carboxyl, alkyl, alkoxy, amino and nitro.In a particular embodiment R₂ is alkyl, cycloalkyl, cycloalkylalkyl,aryl, aralkyl, a heterocycle or heterocyclylalkyl. In a particularembodiment R₂ is alkyl, cycloalkyl or a heterocycle. In a particularembodiment R₂ is selected from the group consisting of t-butyl,isopropyl, cyclohexyl, tetrahydropyran-4-yl,N-methylsulfonylpiperidin-4-yl, tetrahydrothiopyran-4-yl,tetrahydrothiopyran-4-yl (in which the S is in oxidized form SO or SO₂),cyclohexan-4-one, 4-hydroxycyclohexane, 4-hydroxy-4-methylcyclohexane,1-methyl-tetrahydropyran-4-yl, 2-hydroxyprop-2-yl, but-2-yl, phenyl and1-hydoxyeth-1-yl. In an embodiment of the invention R₂ is t-butyl,isopropyl, cyclohexyl, cyclopentyl, phenyl or tetrahydropyran-4-yl. In aparticular embodiment, R₂ is phenyl. In a particular embodiment, R₂ iscyclohexyl. In another embodiment R₂ is tetrahydropyran-4-yl. In anotherparticular embodiment, R₂ is isopropyl (i.e. the valine amino acid sidechain). In another particular embodiment, R₂ is t-butyl. In a particularembodiment R₂ is oriented such that the amino acid, or amino acidanalogue, which it comprises is in the L-configuration.

R₃ is H or alkyl optionally substituted with halogen or hydroxyl; or R₃and R₄ together form a 3-6 heterocycle. In an embodiment R₃ is H oralkyl; or R₃ and R₄ together form a 3-6 heterocycle. In an embodiment R₃is H or methyl, ethyl, propyl or isopropyl. In a particularly particularembodiment R₃ is H or methyl. In a another particular embodiment R₃ ismethyl. In another particular embodiment, R₃ is ethyl. In a particularembodiment R₃ is fluoromethyl. In a particular embodiment R₃ ishydroxyethyl. In another embodiment R₃ is oriented such that the aminoacid, or amino acid analogue, which it comprises is in theL-configuration. In a particular embodiment R₃ and R₄ together with theatoms from which they depend form a 3-6 heterocycle. In a particularembodiment R₃ and R₄ together form an azetidine ring. In a particularembodiment R₃ and R₄ together form a pyrrolidine.

R₃′ is H, or R₃ and R₃′ together form a 3-6 carbocycle. In anembodiment, R₃′ is H. In another embodiment R₃ and R₃′ together form a3-6 carbocycle, for example a cyclopropyl ring. In a particularembodiment R₃ and R₃′ are both methyl.

R₄ and R₄′ are independently H, hydroxyl, amino, alkyl, carbocycle,carbocycloalkyl, carbocycloalkyloxy, carbocycloalkyloxycarbonyl,heterocycle, heterocycloalkyl, heterocycloalkyloxy orheterocycloalkyloxycarbonyl; wherein each alkyl, carbocycloalkyl,carbocycloalkyloxy, carbocycloalkyloxycarbonyl, heterocycle,heterocycloalkyl, heterocycloalkyloxy and heterocycloalkyloxycarbonyl isoptionally substituted with halogen, hydroxyl, mercapto, carboxyl,alkyl, alkoxy, amino, imino and nitro; or R₄ and R₄′ together form aheterocycle. In an embodiment R₄ and R₄′ are independently H, hydroxyl,amino, alkyl, aryl, aralkyl, cycloalkyl, cycloalkylalkyl, heteroaryl, orheteroarylalkyl wherein each alkyl, aryl, aralkyl, cycloalkyl,cycloalkylalkyl, heteroaryl and heteroarylalkyl is optionallysubstituted with halogen, hydroxyl, mercapto, carboxyl, alkyl, alkoxy,amino and nitro; or R₄ and R₄′ together form a heterocycle. In aparticular embodiment R₄ and R₄′ together form a heterocycle, forexample an azetidine ring, or a pyrrolidine ring. In a particularembodiment R₄ and R₄′ are both H. In another particular embodiment R₄ ismethyl and R₄′ is H. In a particular embodiment one of R₄ and R₄′ ishydroxyl (OH) while the other is H. In another embodiment, one of R₄ andR₄′ is amino, such as NH₂, NHMe and NHEt, while the other is H. In aparticular embodiment, R₄′ is H and R₄ is H, alkyl, aryl, aralkyl,cycloalkyl, cycloalkylalkyl, heteroaryl or heteroarylalkyl. In aparticular embodiment R₄ is a group selected from the group consistingof:

R₅ is H or alkyl. In a particular embodiment, R₅ is H or methyl. In aparticular embodiment, R₅ is H. In another particular embodiment, R₅ ismethyl.

R₆, and R₆′ are each independently H, alkyl, aryl or aralkyl. In aparticular embodiment, R₆ is alkyl, for example methyl. In anotherparticular embodiment R₆ is aryl, for example phenyl. In anotherparticular embodiment R₆ is aralkyl, for example benzyl. In a particularembodiment R₆ and R₆′ are the same, for example both alkyl, e.g. bothmethyl. In another particular embodiment R₆ is methyl and R₆′ is H. Inanother embodiment R₆ and R_(6′) are both H.

Compounds of the invention contain one or more asymmetric carbon atoms.Accordingly, the compounds may exist as diastereomers, enantiomers ormixtures thereof. The syntheses of the compounds may employ racemates,diastereomers or enantiomers as starting materials or as intermediates.Diastereomeric compounds may be separated by chromatographic orcrystallization methods. Similarly, enantiomeric mixtures may beseparated using the same techniques or others known in the art. Each ofthe asymmetric carbon atoms may be in the R or S configuration and bothof these configurations are within the scope of the invention. In aparticular embodiment, compounds of the invention have the followingstereochemical configuration of formula I′

wherein ring A, Q, X₁, X₂, Y, R₁, R₂, R₃, R₄′, R₄′, R₅, R₆ and R₆′ areas described herein.

In an embodiment, compounds of the invention have the general formula IV

wherein Q, X₁, X₂, Y, R₁, R₂, R₃, R₄, R₄′, R₅, R₆, R₆ and R₇ are asdescribed here particular embodiment Q is a carbocycle or a heterocycleoptionally substituted with one or more hydroxyl, alkyl, alkoxy, acyl,halogen, mercapto, oxo, carboxyl, amino, cyano, nitro, amidino,guanidino an optionally substituted carbocycle or an optionallysubstituted heterocycle wherein one or more CH₂ or CH groups of saidalkyl is optionally replaced with —O—, —S—, —S(O)—, S(O)₂, —N(R₈)—,—C(O)—, —C(O)—NR₈—, —NR₈—C(O)—, —SO₂—NR₈—, —NR₈—SO₂—, —NR₈—C(O)—NR₈—,—NR₈—C(NH)—NR₈—, —NR₈—C(NH)—, —C(O)—O— or —O—C(O)—. In a particularembodiment, Q is aryl or heteroaryl optionally substituted with one ormore hydroxyl, alkyl, alkoxy, acyl, halogen, mercapto, oxo, carboxyl,amino, cyano, nitro, amidino, guanidino an optionally substitutedcarbocycle or an optionally substituted heterocycle wherein one or moreCH₂ or CH groups of said alkyl is optionally replaced with —O—, —S—,—S(O)—, S(O)₂, —N(R₈)—, —C(O)—, —C(O)—NR₈—, —NR₈—C(O)—, —SO₂—NR₈—,—NR₈—SO₂—, —NR₈—C(O)—NR₈—, —NR₈—C(NH)—NR₈—, —NR₈—C(NH)—, —C(O)—O— or—O—C(O)—. In a particular embodiment Q is aryl or heteroaryl optionallysubstituted with one or more hydroxyl, alkyl, alkoxy, alkoxyalkoxy,acyl, halogen, mercapto, carboxyl, acyl, halo-substituted alkyl, amino,cyano, nitro, amidino, guanidino. In a particular embodiment Q is arylor heteroaryl optionally substituted with halogen, alkyl, alkoxy,alkoxyalkoxy, cyano. In an embodiment Q is IIIa to IIIs wherein R₇, R₈and n are as defined herein. In a particular embodiment Q is IIIq. In aparticular embodiment Q is IIId. In a particular embodiment Q is IIIb,IIIc, IIIe, IIIf, IIIj, IIIk, IIIl, IIIn, IIIo, IIIq, IIIr or IIIs.

In an embodiment when compounds of the invention have the generalformula IV, R₁ is H. In an embodiment when compounds of the inventionhave the general formula IV, R₂ is alkyl, a carbocycle,carbocyclylalkyl, a heterocycle or heterocyclylalkyl each optionallysubstituted with halogen, hydroxyl, mercapto, carboxyl, alkyl, alkoxy,amino and nitro. In an embodiment when compounds of the invention havethe general formula IV, R₃ is H or methyl, ethyl, propyl or isopropyl.In an embodiment when compounds of the invention have the generalformula IV, R₄ is methyl and R₄′ is H. In an embodiment when compoundsof the invention have the general formula IV, R₅ is H. In an embodimentwhen compounds of the invention have the general formula IV, R₆ andR_(6′) are both H. In an embodiment, when compounds of the inventionhave the general formula IV R₇ is H, halogen, cyano, alkyl, hydroxyalkylor alkoxyalkyl. In an embodiment, when compounds of the invention havethe general formula IV, X₁ and X₂ are both O. In an embodiment, whencompounds of the invention have the general formula IV, Y is CH₂.

The invention also encompasses prodrugs of the compounds describedabove. Suitable prodrugs where applicable include known amino-protectingand carboxy-protecting groups which are released, for examplehydrolyzed, to yield the parent compound under physiologic conditions. Aparticular class of prodrugs are compounds in which a nitrogen atom inan amino, amidino, aminoalkyleneamino, iminoalkyleneamino or guanidinogroup is substituted with a hydroxy (OH) group, an alkylcarbonyl (—CO—R)group, an alkoxycarbonyl (—CO—OR), an acyloxyalkyl-alkoxycarbonyl(—CO—O—R—O—CO—R) group where R is a monovalent or divalent group and asdefined above or a group having the formula —C(O)—O—CP1P2-haloalkyl,where P1 and P2 are the same or different and are H, lower alkyl, loweralkoxy, cyano, halo lower alkyl or aryl. In a particular embodiment, thenitrogen atom is one of the nitrogen atoms of the amidino group of thecompounds of the invention. These prodrug compounds are preparedreacting the compounds of the invention described above with anactivated acyl compound to bond a nitrogen atom in the compound of theinvention to the carbonyl of the activated acyl compound. Suitableactivated carbonyl compounds contain a good leaving group bonded to thecarbonyl carbon and include acyl halides, acyl amines, acyl pyridiniumsalts, acyl alkoxides, in particular acyl phenoxides such asp-nitrophenoxy acyl, dinitrophenoxy acyl, fluorophenoxy acyl, anddifluorophenoxy acyl. The reactions are generally exothermic and arecarried out in inert solvents at reduced temperatures such as −78 toabout 50 C. The reactions are usually also carried out in the presenceof an inorganic base such as potassium carbonate or sodium bicarbonate,or an organic base such as an amine, including pyridine, triethylamine,etc. One manner of preparing prodrugs is described in U.S. Ser. No.08/843,369 filed Apr. 15, 1997 (corresponding to PCT publicationWO9846576) the contents of which are incorporated herein by reference intheir entirety.

Particular compounds of formula I include the following:

Synthesis

Compounds of the invention are prepared using standard organic synthetictechniques from commercially available starting materials and reagents.It will be appreciated that synthetic procedures employed in thepreparation of compounds of the invention will depend on the particularsubstituents present in a compound and that various protection anddeprotection steps that are standard in organic synthesis may berequired but may not be illustrated in the following general schemes. Ina particular general synthetic scheme compounds of the invention may beprepared using typical peptide chemistry techniques by coupling aminoacid residue analogues employing typical amide coupling procedures. Inscheme 1, amine-protected amino acid residue analogues are coupled anddeprotected sequentially to give the final compounds.

It will be appreciated that the amino acid analogs may be coupled in anyorder and may be prepared using solid phase support which is routine inthe art. For example, Scheme 2 illustrates an alternative amino acidresidue analogue coupling route.

The intermediate incorporating ring A is prepared from commerciallyavailable reagents employing standard organic chemistry techniques. Forexample, when ring A is thiazole, the intermediate may be preparedaccording to scheme 3.

wherein Q, Y, R₁, R₆, and R₆′ are as defined herein and Pr is an amineprotecting group. A proline analogue wherein the alpha nitrogen isprotected (Pr), for example with Boc or Cbz, and amidated is convertedto the corresponding thioamide, for example using Lawesson's reagentaccording to the procedures described in Williams et al (J. Org. Chem,2001, 66:8463). The thiamide is then cyclized with an appropriatebromide to give the desired thiazole substituted with group Q, forexample using the procedures described in Ciufolini et al, (J. Org.Chem. 1997, 62: 3804). Alternatively, the bromide in the present schememay incorporate a functional group which may be used to couple a desiredgroup Q to the thiazole formed from the cyclization step.

For compounds of the invention in which ring A is an oxazole, theintermediate may be prepared according to scheme 4.

wherein Q, Y, R₁, R₆, and R₆′ are as defined herein and Pr is an amineprotecting group. The starting proline analogue is reacted with anappropriate amine using standard amide forming procedures. The resultingamide is cyclized, for example using Burgess Reagent according to theprocedures described in Pihko et al (J. Org. Chem., 1999, 64:652), togive the dihydro-oxazole. The dihydro-oxazole is then reduced to givethe desired oxazole substituted with group Q. Alternatively, the amineof the first step in the present scheme may incorporate a functionalgroup in place of Q which may be used directly or indirectly to couple adesired group Q to the thiazole formed from the cyclization step.

Compounds of the invention in which R₄ or R₄′ are other than H may beprepared according to standard organic chemistry techniques, for exampleby reductive amination in which a starting amino acid residue analoge.g. NH₂—CH(R₃)—C(O)—OH is reacted with a suitable aldehyde or ketone togive the desired R₄ and R₄′ substituents. See scheme 5. The resultingR₄/R₄′ substituted amino acid intermediate can then be conjugated to thenext amino acid intermediate or the remainder of the compound usingstandard peptide coupling procedures.

In a particular embodiment, alanine is reacted with1-methylindole-2-carboxaldehyde and reduced with sodium cyanoborohydridedissolved in 1% HOAc/DMF to give the N-substituted alanine residue whichmay be used in preparing compounds of the invention. See scheme 6.

Alternatively, the reductive amination procedure to introduce R₄/R₄′substituents is the final step in the preparation of the compound.

When compounds of the invention incorporate R₄ or R₄′ substituents otherthan H, they may also be prepared by substitution of a suitable acidintermediate which incorporates a leaving group with a desired amine.For example Br—CH(R₃)—C(O)—OH is substituted with an amine R₄—NH₂ orR₄—NH—R₄′ according to scheme 7.

Alternatively, the substitution reaction introducing R₄ or R₄′substituents may be performed as a final step in the preparation of thecompound as illustrated in scheme 8.

In a particular embodiment, 2-bromopropionic acid is reacted with thefollowing amines dissolved in DMF and bubbled for until substitution iscomplete to form N-substituted alanine residues:

Compounds of the invention in which either X₁ or X₂ is sulfur, i.e. thecompound incorporates a thioamide, may be prepared according toestablished organic chemistry techniques. For example, compounds inwhich X₂ is sulfur can be prepared according to scheme 9 starting froman Fmoc protected amino acid residue analog NH₂—CH(R₂)—COOH which isdissolved in THF and cooled to −25° C., with addition of DIPEA followedby addition of isobutylchloroformate. After 10 minutes, the diamine,4-nitrobenzene-1,2-diamine, is added and the reaction mixture iscontinuously stirred at −25° C. for 2 hours, then at room temperatureovernight. THF is vacuumed off and the mixture is then subjected toflash chromatography using 50% EtOAc/Hexane to yield the product. TheFmoc-alanine derivative, phosphorus pentasulfide and sodium carbonateare mixed in THF and stirred overnight. The solution is concentrated anddirect chromatography using 80% EtOAc/Hexane yields the activatedthioalanine. The activated thioalanine and sodium nitrite are then mixedin acetic acid and diluted with H₂O. The resulting precipitant isfiltered and dried to yield the product. The thioalanine is coupled toan A ring substituted proline amino acid residue analog by dissolvingboth in DMF. The thioamide product may then be deprotected with 20%PIP/DMA for 15 minutes and used to conjugate to theR₄/R₄′—N—C(R₃)(R₃′)—COOH. Alternatively the Fmoc-protected thioamide isfirst coupled to the A ring substituted proline amino acid residueanalog followed by Fmoc deprotection and subsequent coupling to theR₄/R₄′—R₄/R₄′—N—C(R₃)(R₃′)—COOH amino acid residue analog.

Utility

The compounds of the invention inhibit the binding of IAP proteins tocaspases, in particular X-IAP binding interaction with caspases 3 and 7.The compounds also inhibit the binding of ML-IAP to Smac protein.Accordingly, the compounds of the invention are useful for inducingapoptosis in cells or sensitizing cells to apoptotic signals, inparticular cancer cells. Compounds of the invention are useful forinducing apoptosis in cells that overexpress IAP proteins.Alternatively, compounds of the invention are useful for inducingapoptosis in cells in which the mitochondrial apoptotic pathway isdisrupted such that release of Smac from ML-IAP proteins is inhibited,for example by up regulation of Bcl-2 or down regulation of Bax/Bak.More broadly, the compounds can be used for the treatment of all cancertypes which fail to undergo apoptosis. Examples of such cancer typesinclude neuroblastoma, intestine carcinoma such as rectum carcinoma,colon carcinoma, familiary adenomatous polyposis carcinoma andhereditary non-polyposis colorectal cancer, esophageal carcinoma, labialcarcinoma, larynx carcinoma, hypopharynx carcinoma, tong carcinoma,salivary gland carcinoma, gastric carcinoma, adenocarcinoma, medullarythyroidea carcinoma, papillary thyroidea carcinoma, renal carcinoma,kidney parenchym carcinoma, ovarian carcinoma, cervix carcinoma, uterinecorpus carcinoma, endometrium carcinoma, chorion carcinoma, pancreaticcarcinoma, prostate carcinoma, testis carcinoma, breast carcinoma,urinary carcinoma, melanoma, brain tumors such as glioblastoma,astrocytoma, meningioma, medulloblastoma and peripheral neuroectodermaltumors, Hodgkin lymphoma, non-Hodgkin lymphoma, Burkitt lymphoma, acutelymphatic leukemia (ALL), chronic lymphatic leukemia (CLL), acutemyeloid leukemia (AML), chronic myeloid leukemia (CML), adult T-cellleukemia lymphoma, hepatocellular carcinoma, gall bladder carcinoma,bronchial carcinoma, small cell lung carcinoma, non-small cell lungcarcinoma, multiple myeloma, basalioma, teratoma, retinoblastoma,choroidea melanoma, seminoma, rhabdomyo sarcoma, craniopharyngeoma,osteosarcoma, chondrosarcoma, myosarcoma, liposarcoma, fibrosarcoma,Ewing sarcoma and plasmocytoma.

Compounds of the invention are useful for sensitizing cells to apoptoticsignals. Accordingly, the compounds may be administered prior to,concomitantly with, or following administration of radiation therapy orcytostatic or antineoplastic chemotherapy. Suitable cytostaticchemotherapy compounds include, but are not limited to (i)antimetabolites, such as cytarabine, fludarabine,5-fluoro-2′-deoxyuiridine, gemcitabine, hydroxyurea or methotrexate;(ii) DNA-fragmenting agents, such as bleomycin, (iii) DNA-crosslinkingagents, such as chlorambucil, cisplatin, cyclophosphamide or nitrogenmustard; (iv) intercalating agents such as adriamycin (doxorubicin) ormitoxantrone; (v) protein synthesis inhibitors, such as L-asparaginase,cycloheximide, puromycin or diphtheria toxin; (Vi) topoisomerase Ipoisons, such as camptothecin or topotecan; (vii) topoisomerase IIpoisons, such as etoposide (VP-16) or teniposide; (viii)microtubule-directed agents, such as colcemid, colchicine, paclitaxel,vinblastine or vincristine; (ix) kinase inhibitors such as flavopiridol,staurosporin, ST1571 (CPG 57148B) or UCN-01 (7-hydroxystaurosporine);(x) miscellaneous investigational agents such as thioplatin, PS-341,phenylbutyrate, ET-18-OCH₃, or farnesyl transferase inhibitors(L-739749, L-744832); polyphenols such as quercetin, resveratrol,piceatannol, epigallocatechine gallate, theaflavins, flavanols,procyanidins, betulinic acid and derivatives thereof; (xi) hormones suchas glucocorticoids or fenretinide; (xii) hormone antagonists, such astamoxifen, finasteride or LHRH antagonists. In a particular embodiment,compounds of the present invention are coadministered with a cytostaticcompound selected from the group consisting of cisplatin, doxorubicin,taxol, taxotere and mitomycin C. In a particular embodiment, thecytostatic compound is doxorubicin.

Another class of active compounds which can be used in the presentinvention are those which are able to sensitize for or induce apoptosisby binding to death receptors (“death receptor agonists”). Such agonistsof death receptors include death receptor ligands such as tumor necrosisfactor a (TNF-α), tumor necrosis factor β (TNF-β, lymphotoxin-α), LT-β(lymphotoxin-β), TRAIL (Apo2L, DR4 ligand), CD95 (Fas, APO-1) ligand,TRAMP (DR3, Apo-3) ligand, DR6 ligand as well as fragments andderivatives of any of said ligands. In an embodiment, the death receptorligand is TNF-α. In a particular embodiment, the death receptor ligandis Apo2L/TRAIL. Furthermore, death receptors agonists comprise agonisticantibodies to death receptors such as anti-CD95 antibody, anti-TRAIL-R1(DR4) antibody, anti-TRAIL-R2 (DR5) antibody, anti-TRAIL-R3 antibody,anti-TRAIL-R4 antibody, anti-DR6 antibody, anti-TNF-R1 antibody andanti-TRAMP (DR3) antibody as well as fragments and derivatives of any ofsaid antibodies.

For the purpose of sensitizing cells for apoptosis, the compounds of thepresent invention can be also used in combination with radiationtherapy. The phrase “radiation therapy” refers to the use ofelectromagnetic or particulate radiation in the treatment of neoplasia.Radiation therapy is based on the principle that high-dose radiationdelivered to a target area will result in the death of reproducing cellsin both tumor and normal tissues. The radiation dosage regimen isgenerally defined in terms of radiation absorbed dose (rad), time andfractionation, and must be carefully defined by the oncologist. Theamount of radiation a patient receives will depend on variousconsideration but the two most important considerations are the locationof the tumor in relation to other critical structures or organs of thebody, and the extent to which the tumor has spread. Examples ofradiotherapeutic agents are provided in, but not limited to, radiationtherapy and is known in the art (Hellman, Principles of RadiationTherapy, Cancer, in Principles I and Practice of Oncology, 24875 (Devitaet al., 4th ed., vol 1, 1993). Recent advances in radiation therapyinclude three-dimensional conformal external beam radiation, intensitymodulated radiation therapy (IMRT), stereotactic radiosurgery andbrachytherapy (interstitial radiation therapy), the latter placing thesource of radiation directly into the tumor as implanted “seeds”. Thesenewer treatment modalities deliver greater doses of radiation to thetumor, which accounts for their increased effectiveness when compared tostandard external beam radiation therapy.

Ionizing radiation with beta-emitting radionuclides is considered themost useful for radiotherapeutic applications because of the moderatelinear energy transfer (LET) of the ionizing particle (electron) and itsintermediate range (typically several millimeters in tissue). Gamma raysdeliver dosage at lower levels over much greater distances. Alphaparticles represent the other extreme, they deliver very high LETdosage, but have an extremely limited range and must, therefore, be inintimate contact with the cells of the tissue to be treated. Inaddition, alpha emitters are generally heavy metals, which limits thepossible chemistry and presents undue hazards from leakage ofradionuclide from the area to be treated. Depending on the tumor to betreated all kinds of emitters are conceivable within the scope of thepresent invention.

Furthermore, the present invention encompasses types of non-ionizingradiation like e.g. ultraviolet (UV) radiation, high energy visiblelight, microwave radiation (hyperthermia therapy), infrared (IR)radiation and lasers. In a particular embodiment of the presentinvention UV radiation is applied.

The invention also includes pharmaceutical compositions or medicamentscontaining the compounds of the invention and a therapeutically inertcarrier, diluent or excipient, as well as methods of using the compoundsof the invention to prepare such compositions and medicaments.Typically, the compounds of formula I used in the methods of theinvention are formulated by mixing at ambient temperature at theappropriate pH, and at the desired degree of purity, withphysiologically acceptable carriers, i.e., carriers that are non-toxicto recipients at the dosages and concentrations employed into agalenical administration form. The pH of the formulation depends mainlyon the particular use and the concentration of compound, but may rangeanywhere from about 3 to about 8. Formulation in an acetate buffer at pH5 is a suitable embodiment. In an embodiment, the inhibitory compoundfor use herein is sterile. The compound ordinarily will be stored as asolid composition, although lyophilized formulations or aqueoussolutions are acceptable.

The composition of the invention will be formulated, dosed, andadministered in a fashion consistent with good medical practice. Factorsfor consideration in this context include the particular disorder beingtreated, the particular mammal being treated, the clinical condition ofthe individual patient, the cause of the disorder, the site of deliveryof the agent, the method of administration, the scheduling ofadministration, and other factors known to medical practitioners. The“effective amount” of the compound to be administered will be governedby such considerations, and is the minimum amount necessary to inhibitIAP interaction with caspases, induce apoptosis or sensitize a malignantcell to an apoptotic signal. Such amount is may be below the amount thatis toxic to normal cells, or the mammal as a whole.

Generally, the initial pharmaceutically effective amount of the compoundof the invention administered parenterally per dose will be in the rangeof about 0.01-100 mg/kg, for example about 0.1 to 20 mg/kg of patientbody weight per day, with the typical initial range of compound usedbeing 0.3 to 15 mg/kg/day. Oral unit dosage forms, such as tablets andcapsules, may contain from about 25 to about 1000 mg of the compound ofthe invention.

The compound of the invention may be administered by any suitable means,including oral, topical, transdermal, parenteral, subcutaneous,intraperitoneal, intrapulmonary, and intranasal, and, if desired forlocal treatment, intralesional administration. Parenteral infusionsinclude intramuscular, intravenous, intraarterial, intraperitoneal, orsubcutaneous administration. An example of a suitable oral dosage formis a tablet containing about 25 mg, 50 mg, 100 mg, 250 mg, or 500 mg ofthe compound of the invention compounded with about 90-30 mg anhydrouslactose, about 5-40 mg sodium croscarmellose, about 5-30 mgpolyvinylpyrrolidone (PVP) K30, and about 1-10 mg magnesium stearate.The powdered ingredients are first mixed together and then mixed with asolution of the PVP. The resulting composition can be dried, granulated,mixed with the magnesium stearate and compressed to tablet form usingconventional equipment. An aerosol formulation can be prepared bydissolving the compound, for example 5-400 mg, of the invention in asuitable buffer solution, e.g. a phosphate buffer, adding a tonicifier,e.g. a salt such sodium chloride, if desired. The solution is typicallyfiltered, e.g. using a 0.2 micron filter, to remove impurities andcontaminants.

EXAMPLES

The invention will be more fully understood by reference to thefollowing examples. They should not, however, be construed as limitingthe scope of the invention. Reagents and solvents were obtained fromcommercial sources and used as received. ISCO chromatography refers touse of a pre-packed silica gel columns on a Companion system byTeledyne-Isco, Inc. Lincoln, Nebr. The identity and purity of allcompounds were checked by LCMS and ¹H NMR analysis.

Abbreviations used herein are as follows:

ACN: acetonitrile;

Chg: cyclohexylglycine;

DCM: dichloromethane

DIPEA: diisopropylethylamine;

DMAP: 4-dimethylaminopyridine;

DME: 1,2-dimethoxyethane;

DMF: dimethylformamide;

DMSO: dimethylsulfoxide

EDC: 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide;

EEDQ: 2-ethoxy-1-ethoxycarbonyl-1,2-dihydroquinoline

LCMS: liquid chromatography mass spectrometry;

HATU: O-(7-Azobenzotriazol-1-yl)-1,1,3,3-tetramethyluroniumhexafluorophosphate;

HOBt: N-Hydroxybenzotriazole

HBTU: 2-(1H-Benzotriazol-1-yl)-1,1,3,3-Tetramethyl-uroniumHexafluorophosphate

HPLC: high performance liquid chromatography;

NBS: N-bromosuccinamide;

TASF: tris(dimethylamino)sulfonium difluorotrimethylsilicate;

TEA: triethylamine;

TFA: trifluoroacetate;

THF: tetrahydrofuran;

Example 1 2-[tert-Butoxycarbonyl-(1H-pyrrol-2-ylmethyl)-amino]-propionicacid

Alanine ethyl ester b (5 g, 32.5 mmol), pyrrole-2-carboxaldehyde a (3.1g, 32.5 mmol), sodium cyanoborohydride (2.04 g, 32.5 mmol) and AcOH (1%)were mixed in DMF and stirred overnight. The reaction was quenched withH₂O, and DMF was evaporated. The mixture was diluted with EtOAc, washedby 0.1N NaOH, dried and concentrated to yield product c 2.5 g. Theresulting ester c (2.5 g, 12.8 mmol), di-tert-butyldicarbonate (3.06 g,14 mmol) were mixed in THF, H₂O with NaHCO₃ and stirred overnight. THFwas evaporated, and the mixture was diluted with EtOAc, washed by 1NNaOH, sat. NH₄Cl and brine. After dried, the mixture was concentrated toyield the Boc-protected ester d 3.3 g. The Boc-protected ester d (1.67g, 5.6 mol), lithium hydroxide mono hydrate (284 mg, 6.77 mmol) weremixed in THF and H₂O at 0° C. THF was vacuumed off, and the solution wasacidified by dilute H₂SO₄, extracted by EtOAc twice. Organic layers werecombined, dried and evaporated giving product2-[tert-butoxycarbonyl-(1H-pyrrol-2-ylmethyl)-amino]-propionic acid e.

Example 2 Thiazole Substituted Pyrrolidine

Following the general procedure of Williams (Williams, D. R. et al, M.J. Org. Chem. 2001, 66, 8463), a mixture of N-Cbz-proline amide a (500mg, 2.0 mmol) Lawesson's reagent (420 mg, 1.05 mmol) and toluene (5 mL)was heated at reflux for 2 h. The solution was concentrated, adsorbedonto Celite, and purified by flash chromatography (SiO₂, 40% ethylacetate-hexanes) to afford 393 mg (74%) of compound b as a colorlesssolid.

Following the general procedure of Ciufolini (Ciufolini, M. A. et al, J.Org. Chem. 1997, 62, 3804), ethyl bromopyruvate (200 μl, 1.43 mmol) wasadded to a suspension of thioamide b (378 mg, 1.43 mmol) in ethanol (5mL), and the mixture heated at 80° C. for 5 min. The solvent wasevaporated under reduced pressure, and the residue purified by flashchromatography (SiO₂, gradient elution, 30-40-50% ethyl acetate-hexanes)to afford 393 mg (74%) of thiazole c as a colorless solid.

Phenyl magnesium bromide (2.1 mL of 1.0 M solution in THF, 2.1 mmol) wasadded dropwise to a cold (−78° C.) solution of ester c (360 mg, 1.0mmol) in THF (5 mL) over 5 min. The cooling bath was removed and thesolution allowed to reach room temperature, at which time it was pouredinto saturated aqueous NH₄Cl (50 mL). The aqueous layer was extractedwith 50% ethyl acetate-hexanes (3×10 mL). The combined organic layerswere dried (Na₂SO₄), filtered, and concentrated. The residue waspurified by flash chromatography (SiO₂, gradient elution, 30-40% ethylacetate-hexanes) to afford 404 mg (84%) of thiazole d as a colorlesssolid.

Triethylsilane (850 μl, 5.3 mmol) and TFA (5 mL) were added sequentiallyto alcohol d, and the resulting solution was allowed stand at rt for 1h. The solvent was evaporated, and the residue purified by flashchromatography (SiO₂, 30% ethyl acetate-hexanes) to afford aquantitative yield of compound e as a colorless oil.

Following the general procedure of Thurston (Bose, S. D.; Thurston, D.E. Tetrahedron Lett. 1990, 31, 6903), BF₃.Et₂O (0.78 mL, 6.2 mmol) wasadded to a solution of carbamate e (280 mg, 0.62 mmol), propanethiol(560 μl, 6.2 mmol) and CH₂Cl₂ (3 mL) at rt. After 1 day at rt, thereaction was poured into 1 N NaOH (50 mL) and stirred vigorously for 1h. The layers were separated and the organic phase was washed with 1 NNaOH (2×5 mL). The combined aqueous layers were extracted with CH₂Cl₂(2×5 mL), and the combined organic layers were dried (K₂CO₃), filteredand concentrated. The residue was purified by flash chromatography(SiO₂, gradient elution, 40-50-60% ethyl acetate-hexanes, 1% TEA) toafford 122 mg (61%) of amine f as a colorless solid.

Example 3 Oxazole Substituted Pyrrolidine

A mixture of N-Boc-proline a (5.35 g, 24.9 mmol) serine methyl esterhydrochloride b (3.50 g, 22.5 mmol), EDC (4.76 g, 24.85 mmol), DIPEA(4.0 mL, 22.5 mmol) and CH₂Cl₂ (90 mL) was maintained overnight. Themixture was diluted with CH₂Cl₂ (200 mL) and washed with 1 N HCl (3×100mL), 0.1 N NaOH (3×100 mL) and brine (1×100 mL). The organic layer wasdried (Na₂SO₄), filtered, and concentrated to afford 5.2 g (73%) ofdipeptide c as a colorless foam.

To a cool (0° C.) solution of dipeptide c (4.57 g, 14.4 mmol) and THF(100 mL) was added Burgess Reagent (Pihko, P. M.; Koskinen, A. M. P.;Nissinen, M. J.; Rissanen, K. J. Org. Chem. 1999, 64, 652, andreferences therein) (3.77 g, 15.8 mmol) in 3 portions over 30 min. Thecooling bath was removed and the reaction allowed to reach rt, thenheated at reflux for 1 h. After cooling to rt, the THF was removed underreduced pressure and the residue was partitioned between EtOAc (200 mL)and saturated aqueous NH₄Cl (200 mL). The organic layer was washed withsaturated aqueous NH₄Cl (2×50 mL). The combined aqueous phases wereextracted with EtOAc (1×50 mL) and the combined organic phases werewashed with brine, dried (Na₂SO₄), filtered, and concentrated. Theresidue was purified by flash chromatography (SiO₂, 50-75-100% ethylacetate-hexanes) to afford 2.94 g (68%) of compound d as a colorlesssolid.

Following the general procedure of Koskinen (Pihko, P. M.; Koskinen, A.M. P.; Nissinen, M. J.; Rissanen, K. J. Org. Chem. 1999, 64, 652, andreferences therein), to degassed CH₂Cl₂ (25 mL), was added CuBr (8.79 g,39.3 mmol), hexamethylene tetraamine (5.51 g, 39.3 mmol) and DBU (5.9mL, 39.3 mmol) and the resulting dark mixture stirred vigorously whileit was cooled to 0° C. To this mixture was added a degassed solution ofd (2.94 g, 9.83 mmol) and CH₂Cl₂ (25 mL) over 5 min. The cooling bathwas removed and the mixture stirred vigorously for 2 h. The reaction wasthen poured into 1:1 saturated aqueous NH₄Cl: conc. NH₄OH (200 mL),stirred for 30 min, then extracted with EtOAc (3×50 mL). The combinedorganic phases were washed with saturated aqueous NH₄Cl (2×50 mL),brine, dried (Na₂SO₄), filtered, and concentrated. The residue waspurified by flash chromatography (SiO₂, 40-50% ethyl acetate-hexanes) toafford 1.1 g (38%) of oxazole e as a colorless solid.

Phenylmagnesium bromide (4.4 mL of 1.0 M solution in THF, 4.4 mmol) wasadded dropwise to a cold (−78° C.) solution of ester e (600 mg, 2.0mmol) in THF (10 mL) over 5 min. The cooling bath was removed and thesolution allowed to reach rt, at which time it was poured into saturatedaqueous NH₄Cl (50 mL). The aqueous layer was extracted with 50% ethylacetate-hexanes (3×10 mL). The combined organic layers were dried(Na₂SO₄), filtered, and concentrated. The residue was purified by flashchromatography (SiO₂, gradient elution, 20-30-40% ethyl acetate-hexanes)to afford 443 mg (52%) of oxazole f as a colorless solid.

Triethylsilane (20 μl) and TFA (1 mL) were added sequentially to asolution of alcohol f (50 mg, 0.1 mmol) and CH₂Cl₂ (1 mL). The resultingsolution was allowed stand at rt for 1 h. The solvent was evaporated,and the residue partitioned between EtOAc (20 mL) and 1N NaOH (20 mL).The organic phase was washed with 1N NaOH (2×20 mL). The combinedaqueous phases were extracted with EtOAc (1×20 mL), and the combinedorganic phases were washed with brine (1×20 mL) dried (Na₂SO₄),filtered, and concentrated to afford amine g as a colorless oil,contaminated with residual triethylsilane. This material was useddirectly in the next coupling.

Example 4 Synthesis of Methyl Ketones

A mixture of dihydrobezofuran a (Davies, H. M. L.; Grazini, M. V. A.;Aouad, E. Org. Lett. 2001, 3, 1475) (160 mg, 0.9 mmol) DDQ (300 mg) andCH₂Cl₂ (11 mL) was maintained at room temp. for 2 days. The solution wasdiluted with 50% ethyl acetate-hexanes and washed with 0.5 N NaOH (3×10mL), brine (1×10 mL), dried (Na₂SO₄), filtered, and concentrated toafford 150 mg (93%) of bezofuran b.

Isopropylmagnesium chloride (7.1 mL of a 2.0 M solution in THF, 14.2mmol) was added dropwise to a mixture of benzofuran methyl ester b (500mg, 2.84 mmol) and N, O-dimethyl hydroxyl amine hydrochloride (690 mg,7.1 mmol) and THF (8 mL) maintained ←20° C. The mixture was allowed towarm to 0° C. over 20 min, then poured into 50 mL of saturated aqueousNH₄Cl. The aqueous phase was extracted with EtOAc (3×20 mL), thecombined organic phases were washed with brine (1×50 mL), dried(Na₂SO₄), filtered, and concentrated to afford 577 mg (85%) of amide c,as a clear oil.

To a solution of amide c (660 mg, 3.22 mmol) and THF (6 mL) was addedMeMgBr (3 mL of a 3.0 M solution in THF, 9 mmol) at 0° C. The solutionwas maintained at 0° C. for 30 min, then allowed to warm to 20° C. for30 min, at which time a precipitate forms. The mixture was poured into100 mL of saturated aqueous NH₄Cl. The aqueous phase was extracted withEtOAc (3×50 mL), the combined organic phases were washed with brine(1×50 mL), dried (Na₂SO₄), filtered, and concentrated to afford 460 mg(89%) of ketone d, as a clear oil.

A mixture of potassium tert-butoxide (2.2 g, 17.5 mmol), fluoro ketone e(3.0 g, 15.9 mmol) and ethylene glycol (30 mL) was heated at 50° C. for1 h, then 60° C. for 2 h. The mixture was then poured into 500 mL ofsaturated aqueous NH₄Cl. The aqueous phase was extracted with Et₂O(3×150 mL), the combined organic phases were washed with water (3×150mL), brine (1×50 mL), dried (Na₂SO₄). The mixture was adsorbed ontoCelite, and chromatographed (ISCO, 120 g silica column, 10-60%EtOAc-hexanes) to afford 2.23 g (61%) of the hydroxy ether f as acolorless solid.

A mixture of potassium cyanide (6.9 g, 106 mmol), fluoro ketone e (2.0g, 10.6 mmol) and DMSO (20 mL) was maintained at rt for 4 days, thenheated at 50° C. for 1 day. The mixture was then poured into 500 mL of 1N NaOH. The aqueous phase was extracted with Et₂O (3×150 mL), thecombined organic phases were washed with water (3×150 mL), brine (1×50mL), dried (Na₂SO₄). The mixture was adsorbed onto Celite, andchromatographed (ISCO, 120 g silica column, 0-20% EtOAc-hexanes) toafford 1.15 g (55%) of the nitrile g as a yellow solid.

A mixture of dibromide h (2.33 g, 7.78 mmol), NaSMe (600 mg, 8.56 mmol),and EtOH (5 mL) was maintained at rt 18 h. The mixture was poured into75 mL of 1 N NaOH and extracted with EtOAc (3×50 mL). The combinedorganic phases were washed with 1 N NaOH (1×50 mL), brine (3×50 mL),dried (Na₂SO₄), filtered and concentrated to afford 2.06 g (98%) ofthioether i as a colorless oil.

To a −78° C. solution of bromide i (500 mg, 1.87 mmol) and THF (15 mL)was added sec-BuLi (1.6 mL of 1.4 M solution in cyclohexane, 2.25 mmol)over 5 min. After 5 min at −78° C., the dark purple solution wasquenched rapidly with DMF (0.5 mL) and the solution warmed to 0° C. andmaintained at that temp. for 5 min. The solution was then poured intosaturated aqueous NH₄Cl (50 mL). The aqueous phase was extracted withEtOAc (3×25 mL), the combined organic phases were washed brine (1×50mL), dried (Na₂SO₄) and filtered. The mixture was adsorbed onto Celite,and chromatographed (ISCO, 12 g silica column, 0-10% EtOAc-hexanes) toafford 260 mg (64%) of aldehyde i as a clear oil.

To a solution of aldehyde i (400 mg, 1.86 mmol) and THF (5 mL) was addedMeMgCl (0.9 mL of a 3.0M solution in THF, 2.8 mmol) at 0° C. Thesolution was maintained at 0° C. for 30 min, then allowed to warm to 20°C. for 30 min. The mixture was poured into 50 mL of saturated aqueousNH₄Cl. The aqueous phase was extracted with EtOAc (3×25 mL), thecombined organic phases were washed with brine (1×50 mL), dried(Na₂SO₄), filtered, and concentrated to afford crude alcohol k as aclear oil, which was used without further purification.

To a solution of crude sulfide k and MeOH (5 mL) at 0° C. was added asuspension of Oxone (1.3 g, 2.1 mmol) in water (5 mL) over 20 min. Themixture was allowed to reach room temp and then poured into 50 mL ofsaturated aqueous NH₄Cl. The aqueous phase was extracted with EtOAc(3×25 mL), the combined organic phases were washed with brine (1×50 mL),dried (Na₂SO₄), filtered, and concentrated. This residue was dissolvedin MeOH (10 mL), cooled to 0° C., and to it was added a suspension ofOxone (2.6 g, 4.2 mmol) in water (10 mL) over 20 min. The mixture wasstirred at rt overnight then poured into 50 mL of saturated aqueousNH₄Cl. The aqueous phase was extracted with EtOAc (3×25 mL), thecombined organic phases were washed with brine (1×50 mL), dried(Na₂SO₄), filtered, and concentrated to afford 550 mg (100% for twosteps) of sulfone l as a clear oil.

A mixture of alcohol l (550 mg, 2.1 mmol), Celite (680 mg), and PCC (500mg, 2.31 mmol) was stirred vigorously at rt for 6 h. More PCC (200 mg)was added and the mixture was stirred overnight. The mixture wasadsorbed onto more Celite (5 g) and chromatographed (ISCO, 12 g silicacolumn 0-50% EtOAc-hexanes) to afford 380 mg (69%) of ketone m as acolorless solid.

Thionyl chloride (26 mL, 365 mmol) was added to a mixture of2-methoxy-1-naphthoic acid n (4.5 g, 22.3 mmol) and toluene (45 mL). Theresulting mixture was heated at 75° C. for 3 h. The solvent was removedunder reduced pressure, and the intermediate acid chloride was driedunder high vacuum for 1 h. It was dissolved in THF (50 mL) and cooled to0° C. under N₂. Dimethylzinc (45 mL of 1.0 M solution in heptane, 44.6mmol) was added over 15 min. The reaction mixture was kept at 0° C. for5 min, allowed to warm to room temperature. The reaction was quenchedwith slow addition of saturated NH₄Cl (200 mL). The aqueous phase wasextracted with EtOAc (3×100 mL), and the combined organic phases werewashed with brine (1×100 mL), dried (MgSO₄), filtered, and concentratedin vacuo. The crude product was adsorbed on to Celite and purified byISCO CombiFlash 40 g column (5-15% ethyl acetate-hexane) to afford 1.96g (44%) of ketone o as a white solid.

Following the general procedure of Caldwell (Ichinose, N.; Mizuno, K.;Otsuji, Y.; Caldwell, R. A.; Helms, A. M. J. Org. Chem. 1998, 63,3176-84), to a solution of CH₃MgCl (3.4 mL of 3.0 M solution in THF,10.0 mmol) in THF (20 mL) was added dropwise a solution of4-methoxy-1-naphthalenecarbonitrile p (0.5 g, 2.7 mmol) in toluene (10mL). After the addition, toluene (10 mL) was added to the mixture. Theresulting solution was heated to reflux for 8 h. Aqueous AcOH (50%, 10mL) was added, and the mixture was heated to reflux for 4 h. Aftercooling, the mixture was diluted with water, and the organic phaseseparated, dried (MgSO₄), filtered, and concentrated in vacuo to afford0.5 g (93%) of ketone q as a yellow oil, which was used without furtherpurification.

Following the general procedure of Boswell (Boswell, E. G.; Licause, J.F. J. Org. Chem. 1995, 60, 6592-94), to a solution of sodiumthiomethoxide (0.41 g, 5.8 mmol) in anhydrous DMSO (8 mL) at 0° C. underN₂ was added dropwise a solution of 4-fluoro-1-acetylnaphthalene e (1.0g, 5.3 mmol) in DMSO (8 mL). After stirring at room temperature for 1.5h, the mixture was diluted with water, extracted with CH₂Cl₂ (3×20 mL),and the combined organic phases were dried (MgSO₄), filtered, andconcentrated in vacuo to afford 1.0 g (88%) of sulfide r as a lightyellow solid, which was carried on without further purification.

Following the general procedure of Trost (Trost, B. M.; Curran, D. P.Tetrahedron Lett. 1981, 22, 1287-90), to a cold (0° C.) solution ofsulfide r (2.3 g, 10.6 mmol) in methanol (50 mL) was added dropwise asolution of potassium hydrogen persulfate (Oxone, 22.8 g, 37.1 mmol) inwater (75 mL) keeping the reaction temperature below 5° C. The resultingslurry was stirred at room temperature for 72 h, diluted with water andextracted with CH₂Cl₂ (2×100 mL). The combined organics were washed withbrine, dried (MgSO₄), filtered, and concentrated to afford crudeproduct. The residue was adsorbed on to Celite and purified by ISCOCombiFlash 40 g column (10-40% ethyl acetate-hexane) to afford 2.32 g(88%) of sulfone s as an off white solid.

A mixture of 4-fluoro-1-acetylnaphthalene e (4.75 g, 25.2 mmol),morpholine (6.60 mL, 75.8 mmol), K₂CO₃ (5.21 g, 37.8 mmol), DMSO (30mL), and water (12 mL) was heated at 90° C. for 8 h. The reactionmixture was diluted with water, extracted with CH₂Cl₂ (2×100 mL). Thecombined organic layers were washed with brine, dried (MgSO₄), filtered,and concentrated in vacuo to afford crude product. It was trituratedwith water, filtered, washed with water, dried to afford 6.40 g (99%) ofmorpholinyl ketone t as a yellow solid.

2′-Hydroxy-1′-acetonaphthone u (5.0 g, 26.9 mmol) and K₂CO₃ (11.1 g,81.0 mmol) in acetone (150 mL) were stirred for 20 min. To this mixturewas added bromoethyl methyl ether (3.8 mL, 39.5 mmol) and catalytic KI.The resulting mixture was heated to reflux for 72 h. After cooling, thesolvent was removed in vacuo. The residue was dissolved in EtOAc, washedwith 1 N aqueous NaOH, brine, dried (MgSO₄), filtered, and concentratedin vacuo. The crude product was adsorbed on to Celite and purified byISCO CombiFlash 120 g column (5-25% ethyl acetate-hexane) to afford 3.21g (49%) of ether v as an oil.

Following the general procedure of Short (Short, W. F.; Wang, H. J.Chem. Soc. 1950, 991-4), to a three-necked round-bottomed flask equippedwith a reflux condenser, a dropping funnel, and an aqueous NaOH trap wasadded 1-naphthoic acid w (10.0 g, 58.0 mmol) and AcOH (35 mL). Thissolution was heated at 110° C. and stirred during the addition ofbromine (3.12 mL, 61.0 mmol). After the addition, the mixture was heatedfor another 1.5 h (A yellow solid precipitated during the heating), andthen stirred at room temperature for 24 h. The mixture was poured intoice water. The solid was filtered, washed with water, and crystallizedfrom acetic acid (250 mL) to afford 8.9 g (61%) of bromo acid x as awhite solid.

A solution of bromo acid x (6.0 g, 23.9 mmol), N,O-dimethylhydroxylamine hydrochloride (2.33 g, 23.9 mmol), EDC (4.6 g, 23.9 mmol), andDIPEA (6.3 mL, 35.8 mmol) in DMF (35 mL) was stirred at room temperaturefor 4 h. The mixture was poured into water, extracted with CH₂Cl₂ (2×200mL). The combined organic layers were washed with 0.5 N aqueous HCl, 0.5N aqueous NaOH, dried (MgSO₄), filtered, and concentrated in vacuo. Thecrude product was adsorbed on to Celite and purified by ISCO CombiFlash120 g column (2-10% ethyl acetate-CH₂Cl₂) to afford 4.6 g (65%) of amidey as an oil.

Methylmagnesium chloride (8.5 mL of 3 M solution in THF, 25.5 mmol) wasadded dropwise to a cold (0° C.) solution of amide z (2.5 g, 8.5 mmol)and THF (80 mL). The resulting solution was stirred at 0° C. for 1 h,then allowed to warm to room temperature. After 2.5 h, it was quenchedby slow addition of aqueous AcOH (50%, 10 mL), diluted with water (100mL), and separated. The aqueous layer was extracted with EtOAc (1×100mL). The combined organic layers were washed with brine, dried (MgSO₄),filtered, and concentrated in vacuo to afford 1.9 g (90%) of ketone a′as a yellow solid.

A mixture of 2′-hydroxy-1′-acetonaphthone u (5.0 g, 26.9 mmol), K₂CO₃(7.41 g, 53.7 mmol), and 1-bromo-2-chloroethane (4.4 mL, 53.7 mmol) inDMF (70 mL) was heated at 80° C. for 24 h. The cooled mixture wasdiluted with water, and extracted with CH₂Cl₂ (2×100 mL). The combinedorganic phases were washed with 0.5 N aqueous NaOH, brine, dried(MgSO₄), filtered, and concentrated to afford crude product. The residuewas adsorbed on to Celite and purified by ISCO CombiFlash 40 g column(5-25% ethyl acetate-hexane) to afford 1.6 g (24%) of chloroethoxyketone b′ as a light yellow solid.

A mixture of chloroethoxy ketone b′ (3.0 g, 12.1 mmol), benzoic acid(1.47 g, 12.1 mmol), and Cs₂CO₃ (4.73 g, 14.5 mmol) in DMF (25 mL) washeated at 50° C. for 16 h. Benzoic acid (0.735 g, 6.0 mmol) and Cs₂CO₃(2.36 g, 7.2 mmol) were added, and the mixture heated at 80° C. for 24h. The mixture was filtered, diluted with EtOAc (100 mL), washed withwater, dried (MgSO₄), filtered, and concentrated in vacuo to afford 3.95g (98%) of ketone c′ as a yellow oil.

Following the general procedure of Oda (Oda, M.; Yamamuro, A.; Watabe,T. Chem. Lett. 1979, 1427-30), Trimethylsilyl cyanide (4.5 mL, 34.1mmol) was added slowly into a mixture of 5-methoxy-1-tetralone d′ (5.0g, 28.4 mmol), catalytic ZnI₂ in toluene (12 mL). The resulting mixturewas stirred at room temperature for 24 h. Pyridine (40 mL) and POCl₃(8.0 mL, 85.2 mmol) were added, and the mixture was heated to reflux for8 h. The cooled dark solution was poured into ice water (300 mL) andconc. HCl (10 mL) with stirring, extracted with EtOAc (3×400 mL). Thecombined organic layers were washed with brine, dried (MgSO₄), filtered,and concentrated to afford 4.78 g of crude unsaturated nitrile e′ as abrown solid.

A mixture of the above unsaturated nitrile e′ (4.78 g, 25.8 mmol) andDDQ (5.86 g, 25.8 mmol) in toluene (100 mL) was heated at 100° C. for3.5 h. After cooling, the precipitate was removed by filtration, andwashed with toluene. The combined toluene layers were washed with 0.5 NNaOH (2×100 mL), dried (MgSO₄), and concentrated in vacuo to afford 4.22g (81%) of nitrile f′ as a yellow solid, which was carried on withoutfurther purification.

Following the general procedure for conversion of p to q, nitrile f′(2.20 g, 12.0 mmol) afforded 1.64 g (68%) of ketone g′ as a brown oil.

(2-Chloroethoxy)trimethylsilane (8.70 mL, 53.8 mmol) was added to themixture of 2′-hydroxy-1′-acetonaphthone u (5.0 g, 26.9 mmol), KOH (3.0g, 53.8 mmol) in DMSO (60 mL) and water (20 mL). The resulting mixturewas heated at 80° C. for 24 h. The mixture was diluted with water (400mL). The crystalline precipitate was collected by filtration, washedwith water, dried to afford 5.21 g (84%) of hydroxy ketone h′ as a brownsolid.

Bromine (610 μl, 11.9 mmol) was added over 10 min to a solution ofhydroxy ketone h′ (2.50 g, 10.9 mmol) in CH₂Cl₂ (30 mL) and AcOH (8.0mL) at room temperature. After 2 h, it was quenched with 10% aqueousNa₂S₂O₃ (5 mL), diluted with CH₂Cl₂ (50 mL). The layers were separatedand the aqueous layer was extracted with 50 mL of CH₂Cl₂. The combinedorganics were washed with 0.5 N aqueous NaOH until the aqueous washesare basic, dried (MgSO₄), filtered, and concentrated to afford 3.70 g(96%) of bromo ketone i′ as a dark brown oil.

A mixture of 2-methoxyethanol (3.35 mL, 42.5 mmol) and potassiumt-butoxide (4.76 g, 42.5 mmol) in THF (80 mL) was stirred at roomtemperature for 10 min. To this mixture was added dropwise a solution of4-fluoro-1-acetylnaphthalene (4.0 g, 21.3 mmol) in THF (20 mL), and themixture was stirred at room temperature for 24 h. The mixture wasdiluted with water (50 mL), and the phases separated. The organic layerwas washed with 0.5 N NaOH, brine, dried (MgSO₄), filtered, andconcentrated to afford 5.6 g (106%, excess wt. is solvent) of ketone j′as a brown liquid which solidified under high vacuum.

Following the general procedure of Tagat (Tagat, J. R.; McCombie, S. W.;Nazareno, D. V.; Boyle, C. D.; Kozlowski, J. A.; Chackalamannil, S.;Josien, H.; Wang, Y.; Zhou, G. J. Org. Chem. 2002, 67, 1171-77), asuspension of the bromo acid x (3.0 g, 12.0 mmol) in toluene (18 mL) washeated at 80° C. To this reaction mixture was added dropwiseN,N-dimethylformamide di-tert-butyl acetal (10.0 mL, 42 mmol), and theresulting mixture was heated for an additional 30 min. It was cooled tort, washed with water, saturated aqueous NaHCO₃, brine, dried (Na₂SO₄),filtered, and concentrated in vacuo to afford 2.87 g (78%) of t-butylester k′ as a yellow oil, which was carried on without furtherpurification.

Following the general procedure of Tagat, a stirred solution of t-butylester k′ (1.4 g, 4.5 mmol) in anhydrous THF (30 mL) was cooled to −78°C. under N₂. n-BuLi (3.65 mL of 1.6 M solution in hexane, 5.85 mmol) wasadded, and the resulting solution was stirred for 2 min, followed byaddition of a solution of N-fluorobenzenesulfonimide (2.83 g, 9.0 mmol)in THF (10 mL). After stirring at −78° C. for 30 min, the reaction wasquenched at −78° C. with saturated aqueous NH₄Cl. The aqueous layer wasextracted with Et₂O (2×50 mL), dried (MgSO₄), filtered, and concentratedin vacuo. The crude material was adsorbed on to Celite and purified byISCO CombiFlash 40 g column (2-20%, EtOAc-hexane) to afford 0.57 g (52%)of fluoro compound l′ as colorless liquid.

Trifluoroacetic acid (3.85 mL, 50 mmol) was added to a stirred solutionof fluoro compound l′ (1.23 g, 5.0 mmol) in CH₂Cl₂ (50 mL) at rt. Afterstirring for 3 h, the solution was concentrated in vacuo to afford 0.95g (100%) of fluoro acid m′ as an oil, which was carried on.

A mixture of fluoro acid m′ (820 mg, 4.3 mmol), N, O-dimethylhydroxylamine hydrochloride (420 mg, 4.3 mmol), EDC (825 mg, 4.3 mmol), andDIPEA (750 μl, 4.3 mmol) in DMF (12 mL) was stirred at rt for 3 h. Themixture was diluted with EtOAc (50 mL), washed with 10% citric acid, 0.5N NaOH, dried (MgSO₄), filtered, adsorbed on to Celite, and purified byISCO CombiFlash 12 g column (2-10%, EtOAc-hexane) to afford 0.48 g (48%)of fluoro amide n′ as an oil.

To a solution of fluoro amide n′ (1.07 g, 4.6 mmol) in THF at 0° C. wasadded dropwise a solution of CH₃MgCl (4.6 mL of 3 M solution in THF,13.8 mmol). The resulting mixture was stirred at 0° C. for 1 h, then 2 hat rt. The mixture was quenched with 50% aqueous AcOH (10 mL), dilutedwith water (50 mL), EtOAc (50 mL), and separated. The aqueous layer wasextracted with EtOAc (50 mL). The combined EtOAc layers were dried(MgSO₄), filtered, and concentrated to afford 0.77 g (89%) of fluoroketone o′ as an oil.

Following the general procedure of Coudret (Hortholary, C.; Coudret, C.J. Org. Chem. 2003, 68, 2167-74), to a solution of4-amino-1-naphthalenecarbonitrile p′ (5.0 g, 29.7 mmol) in conc. HCl (50mL) at 0° C. was carefully added sodium nitrite (3.07 g, 44.5 mmol). Themixture was stirred at 0° C. for 1 h, then transferred into anadditional funnel, and added dropwise to an ice-cold solution of CuCl(5.3 g, 53.5 mmol) in water (150 mL). After addition, CH₂Cl₂ (80 mL) wasadded to the reaction mixture. The resulting mixture was allowed to warmto rt and was stirred for 4 h. The mixture was diluted with CH₂Cl₂, andthe phases separated. The aqueous phase was carefully extracted withCH₂Cl₂ (2×150 mL). The combined CH₂Cl₂ phases were washed once withsaturated sodium thiosulfate, dried (MgSO₄), filtered, adsorbed on toCelite, and purified by ISCO CombiFlash 120 g column (2-12%,EtOAc-hexane) to afford 2.63 g (46%) of chloro compound q′ as whitesolid.

Following the general procedure for conversion of p to q, chlorocompound q′ (2.63 g, 14.1 mmol) afforded 2.1 g (74%) of chloro ketone r′as a yellow liquid.

Following the procedure of Hallberg (Alterman, M.; Hallberg, A. J. Org.Chem. 2000, 68, 7984-89) a mixture of bromo ketone x (1.40 g, 5.62mmol), Zn(CN)₂ (790 mg, 6.74 mmol), Pd(PPh₃)₄ (216 mg, 0.19 mmol) andDMF (8 mL) was heated in a microwave reactor (Emry's Optimizer) in asealed heavy-walled tube at 180° C. for 5 min. After cooling, it wasdiluted with water (30 mL), extracted with EtOAc (50 mL), dried (MgSO₄),filtered, adsorbed on to Celite, and purified by ISCO CombiFlash 40 gcolumn (5-20%, EtOAc-hexane) to afford 900 mg (83%) of nitrile ketone s′as white solid.

Following the procedure of Leadbeater (Arvela, R.; Leadbeater, N. E.SynLett. 2003, 8, 1145-48), a mixture of bromo ketone x (100 mg, 0.40mmol), NiCl₂ (103 mg, 0.80 mmol) and DMF (2 mL) was heated in amicrowave reactor (Emry's Optimizer) in a sealed heavy-walled tube at200° C. for 8 min. After cooling, it was diluted with water (15 mL),extracted with EtOAc (20 mL), dried (MgSO₄), filtered, adsorbed on toCelite, and purified by ISCO CombiFlash 4 g column (5-15%, EtOAc-hexane)to afford 55 mg (68%) of chloro ketone t′ as off white solid.

Example 5 Bromination of Methyl Ketones and Preparation of Thiazoles

Bromine (260 μl, 5.07 mmol), was added over 20 min to a solution ofketone a (784 mg, 4.6 mmol) in CH₂Cl₂ (10 mL). The solution wasmaintained at rt for 1 h, then quenched with 10% aqueous Na₂S₂O₃ (10 mL)and stirred vigorously for 20 min. The layers were separated and theorganic phase washed with saturated aqueous NaHCO₃ (1×10 mL), brine(1×10 mL), dried (Na₂SO₄), filtered, and concentrated to afford 1.15 gof bromo ketone b as a yellow oil. Analysis by ¹H NMR indicates a70:15:15 mixture of product to starting ketone and dibrominatedmaterial.

A particular procedure: A mixture of Boc-proline-amide c (8.4 g, 39.2mmol), Lawesson's reagent (8.25 g, 20.4 mmol) and toluene was heated at50° C. for 1 h (use of higher temperatures results in loss ofenantiopurity). The mixture was then adsorbed onto Celite, and purifiedby chromatography (ISCO, 120 g silica column, gradient elution 10-70%EtOAc-hexanes) to afford 7.6 g (84%) of the thioamide d as a colorlesssolid.

A particular procedure for thiazole formation: A mixture of thioamide d(7.81 g, 34 mmol), bromoketone b (7.05 g, 80% pure by ¹H NMR, 22.6mmol), pyridine (1.76 mL, 20.3 mmol) and ethanol (75 mL) was heated at80° C. for 1 h. The ethanol was removed under reduced pressure, and theresidue was adsorbed onto Celite. The residue was chromatographed (SiO₂,gradient elution 0-2.5-5% EtOAc/CH₂Cl₂) to afford 6.3 g (73%) ofthiazole e as a colorless solid.

A mixture of bromide f (145 mg, 0.33 mmol), PhB(OH)₂ (107 mg, 0.88mmol), K₂CO₃ (825 μl of 2.0 M aqueous solution, 1.65 mmol), Pd(PPh₃)₄(15 mg, 0.13 mmol), and 20% EtOH-toluene (2.5 mL) was maintained at 80°C. for 3 h. The mixture was diluted with CH₂Cl₂ (10 mL), and washed with1 N NaOH (2×5 mL). The combined aqueous layers were extracted withCH₂Cl₂ (1×10 mL). The combined organic phases were washed with brine(1×10 mL), dried (Na₂SO₄), filtered, adsorbed on to Celite, and purifiedby flash chromatography (SiO₂, 10-15-20% acetone-hexanes) to afford 74mg (52%) of thiazole g as a colorless solid.

Thiazole e (70 mg, 0.18 mmol) in 1:1 dichloromethane:hexanes (1.5 mL),was treated with N-chlorosuccinimide (30 mg 0.22 mmol). The reactionmixture was stirred at rt for 2 h, at which point additional NCS (10 mg)was added and the mixture stirred overnight. Celite was added, and thedichloromethane was removed under reduced pressure. The product waspurified by chromatorgraphy (ISCO, 12 g silica column, gradient elution0-30% EtOAc/hexanes) to afford 70 mg (99%) of chlorothiazole h.

Thiazole e (120 mg, 0.31 mmol) in dichloromethane (1.5 mL), was treatedwith N-bromosuccinimide (65 mg 0.37 mmol). The reaction mixture wasstirred at room temperature for 3 h. After this period, Celite wasadded, and the dichloromethane was removed under reduced pressure. Theproduct was purified by chromatorgraphy (ISCO, 12 g silica column,column was first flushed with CH₂Cl₂ for 7 minutes and then a gradientof 0-9% EtOAc/CH₂Cl₂ gradient over 9 minutes.) to afford 128 mg (90%) ofbromide i.

Following literature precedent ((1) Maguire, M. P.; Sheets, K. R.;McVety, K.; Spada, A. P.; Zilberstein, A. J. Med. Chem. 1994, 37,2129-2137; (2) Moreno, I.; Tellitu, I.; Dominguez, E.; SanMartin, R.;Eur. J. Org. Chem. 2002, 2126-2135) a mixture of bromothiazole i, (280mg, 0.61 mmol) and alkynylstannane j (Dabdoub, M. J.; Dabdoub, V. B.;Baroni, A. C. M. J. Am. Chem. Soc. 2001, 123, 9694-9695) (250 mg, 0.73mmol), LiCl (approximately 50 mg, 120 mmol) and toluene (6 mL) wasdegassed with nitrogen for 30 min.Tetrakis(triphenylphosphine)palladium(0) (28 mg, 0.02 mmol), was addedand the mixture was heated at 100° C. 3 h. After cooling, Celite wasadded to the mixture, and the solvents were removed under reducedpressure. The residue was purified by chromatography (ISCO, 12 g silicacolumn, column was first flushed with CH₂Cl₂ for 5 minutes and then agradient of 0-20% EtOAc/CH₂Cl₂ gradient over 10 minutes.) to afford 160mg (60%) of alcohol k.

Following literature precedent (Neidlein, R.; Nussbaumer, THeterocycles, 2000, 52, 349), bromide i (600 mg, 1.3 mmol),TMS-acetylene l (1.8 mL, 13 mmol) and TMG (0.6 mL, 5 mmol), weredissolved in dimethylacetamide (6 mL). This mixture was degassed withnitrogen for 30 min. Bis(triphenylphosphine)palladium dichloride (46 mg,0.07 mmol) and copper(I) iodide (62 mg, 0.3 mmol) were added and themixture was sealed and heated at 70° C. for 30 minutes. The mixture wasdiluted with ½-saturated ammonium chloride and filtered through a pad ofcelite. The aqueous mixture was extracted with 70% diethyl ether inhexane (3×20 mL), dried (Na₂SO₄), filtered, adsorbed on to Celite, andchromatographed (ISCO, 40 g column and a solvent gradient of 0-11% ethylacetate in hexane after flushing with hexane for 3 minutes). Terminalalkyne product 27 mg (5%) was isolated along with 200 mg of silylderivative. The TMS group was removed from this material by treatmentwith potassium carbonate (200 mg) in methanol (5 mL) for 3 hours at rt.Celite and toluene (1 mL) were added to the mixture, and the solventswere removed under reduced pressure. The product was purified bychromatography (ISCO 40 g column, solvent gradient of 0-11% ethylacetate/hexane after flushing with pure hexane for 3 minutes), to afforda further 110 mg of terminal alkyne m (26% combined).

Terminal alkyne m (50 mg, 0.12 mmol) was dissolved in THF (0.3 mL) andcooled to −78° C. LHMDS (0.15 mL of a 1.0 M solution of in THF, 0.15mmol) was added dropwise and allowed to stir for 10 minutes. Methyliodide (0.1 mL, excess) was added, the reaction was stirred for 10minutes at −78° C. and then allowed to gradually warm to rt, over 45minutes. Celite was then added to the reaction mixture, the solventswere evaporated under reduced pressure, and the residue purified bychromatography (ISCO, 12 g, column gradient elution 0-18% ethyl acetatein hexane) to afford 25 mg (63%) of the methyl alkyne n.

Typical Boc deprotection: Carbamate o (75 mg, 0.18 mmol) was treatedwith TFA (2 mL) and water (2 drops), in CH₂Cl₂ (2 mL) for 2 h. Thevolatiles were removed under reduced pressure, the residue dissolved inethyl acetate (10 mL) and washed with 1 N NaOH (3×3 mL). The combinedaqueous layers were extracted with ethyl acetate (1×2 mL). The combinedorganic phases were washed with brine (1×3 mL), dried (Na₂SO₄),filtered, and concentrated to provide quantitative yield of amine p.

Example 6 Linear Coupling Procedure

Typical HATU coupling: A mixture of amine a (169 mg, 0.59 mmol),N-Boc-t-butly glycine (150 mg, 0.65 mmol), HATU (450 mg, 1.18 mmol),DIPEA (200 μl, 1.18 mmol) and DMF (2 mL) was maintained at rt for 2 h.The solution was diluted with ethyl acetate (50 mL) and washed with 1 NHCl (3×10 mL), 1 N NaOH (3×5 mL), brine (1×10 mL), dried (Na₂SO₄),filtered, and concentrated. The residue was purified by flashchromatography (SiO₂, 10-15-20% ethyl acetate-hexanes) to afford 286 mg(97%) of amide b as a colorless solid.

Following the general Boc deprotection procedure described above, Bocamine b (317 mg, 0.64 mmol) afforded a quantitative yield of amine c asa colorless solid.

Typical EDC coupling: A solution of amine c (300 mg, 0.76 mmol),N-Boc-alanine (158 mg, 0.84 mmol), EDC (161 mg, 0.84 mmol), catalyticDMAP and MeCN (3 mL) was maintained at rt for 3 h. The solution wasdiluted with ethyl acetate (50 mL) and washed with 1 N HCl (3×10 mL), 1N NaOH (3×5 mL), brine (1×10 mL), dried (Na₂SO₄), filtered, andconcentrated to provide 453 mg of crude residue d, which was carried ondirectly:

Typical final Boc removal and purification: The crude residue d fromabove was treated with TFA (2 mL) and water (2 drops), in CH₂Cl₂ (2 mL)for 2 h. The volatiles were removed under reduced pressure. The residuewas purified by reverse-phase HPLC (C₁₈, MeCN—H₂O, 0.1% TFA) and thesolvents removed by lyophylization to provide 166 mg (38% for 2 steps)of amine e as a colorless powder.

Example 7 N-Boc-N-methyl-L-alanine-L-cyclohexylglycine

A solution of Fmoc-L-cyclohexylglycine (3.6 g, 9.6 mmol) dissolved inDCM (50 mL) and DIPEA (5.6 mL, 32 mmol) was added to 2-chlorotritylchloride resin (5 g, 8 mmol) and gently agitated for 3 hours at roomtemperature. The resin was washed with DCM 4 times, DCM/MeOH/DIPEA(17:2:1) 3 times, DCM 3 times, and 2 times dimethylacetamide (DMA). TheFmoc group was removed by treating the resin with 20% piperidine/DMA (50mL) for 15 minutes. The resin was washed with DMA 6 times. A solution ofBoc-N-methylalanine (3.3 g, 16 mmol), HBTU (6.1 g, 16 mmol), and DIPEA(5.6 mL, 32 mmol) and DMA/DCM (1: 1, 50 mL) was added to the resin andgently agitated for 2 hours at room temperature. The resin was washedwith DMA 5 times, DCM 2 times, and dried under reduced pressure. Thedipeptide was cleaved from the resin by gentle agitation withHOAc/TFE/DCM (1:1:3, 100 mL) for 2 hours at room temperature. The resinwas removed by filtration and the solution concentrated. Residual AcOHwas removed by azeotroping with hexanes (15 times volume). The solidresidue was purified by reverse-phase HPLC (C₁₈, MeCN—H₂O, 0.1% TFA) andthe solvents removed by lyophylization to provide 1.2 g (43%) ofdipeptide N-Boc-N-methyl-L-alanine-L-cyclohexylglycine as a whitepowder.

Example 8 N-Boc-N-methyl-L-alanine-L-dehydropyranylglycine

A mixture of N-Cbz-dehydropyranylglycine methyl ester a (Burk, M. J.;Gross, M. F.; Martinez, J. P. J. Am Chem. Soc. 1995, 117, 9375, andreferences therein) (5.2 g, 17 mmol), 5% Pd.C (500 mg), MeOH (75 mL) andTHF (25 mL) was maintained under an atmosphere of H₂ for 24 h. Themixture was filtered through Celite and the Celite washed with MeOH, andconcentrated under reduced pressure to afford a quantitative yield ofamine b as a colorless oil, which was carried on directly.

The amine b prepared above was combined with CH₂Cl₂ (40 mL), saturatedaqueous NaHCO₃ (40 mL) and cooled to 0° C. Benzyloxy carbonyl chloride(3.0 mL) was then added dropwise and the mixture stirred vigorouslyovernight. The phases were separated and the aqueous phase extractedwith CH₂Cl₂ (3×20 mL). The combined organic phases were washed withbrine (1×50 mL), dried (Na₂SO₄), filtered, adsorbed onto Celite andchromatographed (ISCO, 120 g silica column, gradient elution 5-55%EtOAc-hexanes) to afford 4.15 g (80%) of racemic Cbz-pyranylglycinemethyl ester. The enantiomers were separated on a Chiracel OD columneluting with 10% EtOH-hexanes. The desired S-enantiomer c elutes firstunder these conditions.

A mixture of (S)—N-Cbz-pyranyl glycine c methyl ester (2.4 g, 7.82 mmol)10% Pd.C (700 mg), MeOH (80 mL) was maintained under 1 atmosphere of H₂for 24 h. The mixture was filtered through Celite with MeOH, andconcentrated under reduced pressure to afford 1.35 g (100%) of amine das a colorless oil. Alternatively, pyranyl glycine can be synthesized inenantiopure form following the procedure of Ghosh (Ghosh, A. K.;Thompson, W. J.; Holloway, M. K.; McKee, S. P.; Duong, T. T.; Lee, H.Y.; Munson, P. M.; Smith, A. M.; Wai, J. M.; Darke, P. L.; Zugay, J. A.;Imini, E. A.; Schleif, W. A.; Huff, J. R.; Anderson, P. S. J. Med.Chem., 1993, 36, 2300).

A mixture of amine d (1.35 g, 7.8 mmol), N-Boc-N-methyl alanine e (1.74g, 8.6 mmol), EDC (1.65 g 8.8 mmol) and MeCN (50 mL) was maintained atrt overnight. The MeCN was removed under reduced pressure, and theresidue diluted with EtOAc, washed with 0.5 N HCl (3×10 mL), 0.5 N NaOH(3×10 mL), dried (MgSO₄), filtered, and concentrated to provide 2.1 g(75%) of protected dipeptide f, as a clear oil.

To a 0° C. solution of ester f (2.10 g, 5.86 mmol) and THF (50 mL) wereadded LiOH.H₂O (1.23 g, 29.3 mmol) and water (2 mL). The mixture wasmaintained at 0° C. for 2 h, then the cooling bath was removed and themixture was stirred overnight. Most of the THF was then removed underreduced pressure and the residue was diluted with CH₂Cl₂, washed with0.5 N HCl, dried (MgSO₄), filtered, and concentrated to provide 1.53 g(78%) of dipeptide N-Boc-N-methyl-L-alanine-L-dehydropyranylglycine g,as a colorless solid.

A particular procedure for convergent coupling: A mixture of amine h (69mg, 0.26 mmol), dipeptide N-Boc-N-methyl-L-alanine-L-cyclohexylglycinefrom example 7 (60 mg, 0.23 mmol), HOAt (Carpino, L. A.; El-Faham, A.Tetrahedron, 1999, 55, 6813) (47 mg, 0.24 mmol), DIC (53 μl, 0.34 mmol)and CH₂Cl₂ (2 mL) was maintained at rt overnight. The mixture wasadsorbed onto Celite and purified by chromatography (ISCO, 4 g silicacolumn, gradient elution 5-50% EtOAc-hexanes) to afford 94 mg of theproduct i as a colorless solid contaminated with diisopropyl urea. Themixture was carried on directly to the next step.

The crude residue i from above was treated with TFA (2 mL) and water (2drops), in CH₂Cl₂ (2 mL) for 2 h. The volatiles were removed underreduced pressure. The residue was purified by reverse-phase HPLC (C₁₈,MeCN-—₂O, 0.1% TFA) and the solvents removed by lyophylization toprovide 77 mg (54% for 2 steps) of amine salt j as a colorless powder.

A mixture of the Acetate product k (228 mg, 0.32 mmol), K₂CO₃ (53 mg,0.38 mmol) in aqueous methanol (1:2, v:v, 15 mL) was stirred at rt for 1h. Methanol was removed in vacuo. The residue was diluted with water,extracted with CH₂Cl₂ (1×50 mL), and the organic phase dried (MgSO₄),and concentrated in vacuo to afford a crude product. Conversion to theamine salt I was accomplished in 18% yield (3 steps) following thegeneral procedure.

Example 8

A mixture of acid a (1.5 g, 6.9 mmol) prepared according to theprocedures of described in Kice et al. (J. Org. Chem. 1989, 54,3596-3602), amine HCl salt (868 mg, 8.9 mmol), EDC (1.3 g, 6.9 mmol),and DIPEA (1.2 mL, 6.9 mmol) in DMF (17 mL) was stirred at RT forovernight. The mixture was diluted with EtOAc (50 mL), washed with 0.5 NHCl, 0.5 N NaOH, dried (MgSO₄), filtered, concentrated in vacuo toafford 1.3 g (74%) of amide b as a yellow solid, which was carried onwithout further purification.

10% Pd/C (200 mg) was added into a solution of acid a (500 mg, 2.3mmol), NaOH (92 mg, 2.3 mmol) in EtOH (25 mL) and water (5 mL) in a Parrreactor. This mixture was purged with N₂ for 10 min, then hydrogenatedwith a Parr hydrogenator at 50 psi at RT for 2.5 h. The resultingmixture was filtered through Celite, concentrated in vacuo to afford 50mg (104%) of amine salt b as a greenish-brown solid.

Example 10

To a stirred mixture of the amine salt a (500 mg, 2.4 mmol) in 6N HCl(30 mL), cooled at 0° C., was added NaNO₂ (248 mg, 3.6 mmol) in oneportion (caution was used due to elevated reaction temperature). Afterstirred at 0° C. for 1 h, this solution was added drop wise, via adropping funnel, over 20 min to an ice-water cold solution of CuBr (618mg, 4.3 mmol) in water (30 mL). Then dichloromethane (40 mL) was addedto slowly to the reaction mixture (caution was used due to foaming). Theresulting mixture was allowed to reach RT and was stirred for 4 h. Itwas diluted with CH₂Cl₂ (100 mL), separated, washed the aqueous layerwith another portion of CH₂Cl₂ (100 mL). The combined CH₂Cl₂ were washedonce with sat. aq. Na₂S₂O₃, dried (MgSO₄), and concentrated in vacuo.The crude product was adsorbed on to Celite and purified by ISCOCombiFlash 12 g column (20-100% ethyl acetate-hexane) to afford 175 mg(29%) of bromo acid b as a light yellow solid.

Example 11

A mixture of bromo acid a (680 mg, 2.7 mmol), amine HCl salt (264 mg,2.7 mmol), EDC (520 mg, 2.7 mmol), and DIPEA (472 μL, 2.7 mmol) in DMF(10 mL) was stirred at RT for overnight. The mixture was partitionbetween water (50 mL) and EtOAc (100 mL), separated, washed the aqueouslayer with another portion of EtOAc (100 mL). The combined organic werewashed with 1N HCl (50 mL), 1N NaOH (50 mL), dried (MgSO₄), filtered,concentrated in vacuo. The crude product was adsorbed on to Celite andpurified by ISCO CombiFlash 12 g column (10-50% ethyl acetate-hexane) toafford 300 mg (38%) of bromo amide b as a yellow oil.

Example 12

To a solution of bromo amide a (300 mg, 1.0 mmol) in THF (8 mL) at 0° C.was added drop wise a solution of CH₃MgCl (2.0 mL of 3 M solution inTHF, 6.0 mmol). The resulting mixture was stirred for 1 h, and thenallowed to warm to RT for 2 h. The mixture was quenched with 50% aq.AcOH (4 mL), diluted with water (50 mL) and EtOAc (50 mL), separated.The aqueous layer was extracted with EtOAc (50 mL). The combined EtOAcwere dried (MgSO₄), filtered, and concentrated. The crude product wasadsorbed on to Celite and purified by ISCO CombiFlash 12 g column (2-10%ethyl acetate-hexane) to afford 150 mg (60%) of bromo ketone b as lightyellow oil. Compound b may also be prepared employing the proceduresdescribed by Alvaro et al. (WO 2004099143) and Tsuno et al. (Bull Chem.Soc. of Japan 1975, 48(11), 3347-55).

Example 13

The starting acid a (6.6 g, 30.4 mmol) was treated with thionyl chloride(50 mL), CHCl₃ (50 mL), and 1 drop of DMF at 78° C. for 5 h. Theresulting mixture was concentrated in vacuo, dried under high vacuum forovernight. The resulting yellow solid was cooled in an ice-water bathand MeOH (200 mL) was added slowly. This was then refluxed for 1 h.After cooled to RT, the resulting precipitate was collected byfiltration, washed with cold MeOH, and dried to afford 5.0 g (71%) ofester b as a yellow solid.

Example 14

A suspension of ester a (5.0 g, 21.6 mmol) and 10% Pd/C (1.2 g) in MeOH(200 mL) was purged with N₂ for 5 min, then treated with a balloon of H₂at RT until the reaction is completed (checked by LCMS). After purgingthe reaction mixture with N₂ for 10 min, the mixture was filteredthrough Celite, washed with MeOH, concentrated in vacuo, and high vacuumdried to afford 4.2 g (97%) of amine b as a brown oil.

Example 15

A mixture of bromo ketone a (285 mg, 1.1 mmol), Zn(CN)₂ (400 mg, 3.4mmol), and Pd(PPh₃)₄ (88 mg, 0.076 mmol) in DMF (2 mL) was heated in amicrowave at 200° C. for 600 sec. After cooled, diluted with water (10mL), extracted with EtOAc (2×10 mL). The insoluble material was removedby filtration; the solvent was dried (MgSO₄), and concentrated. Thecrude product was adsorbed on to Celite and purified by ISCO CombiFlash12 g column (5-20% ethyl acetate-hexane) to afford 147 mg (66%) ofnitrile ketone b as a white solid. Compound a may also be preparedemploying the procedures described by Alvaro et al. (WO 2004099143) andTsuno et al. (Bull Chem. Soc. of Japan 1975, 48(11), 3347-55).

Example 16

To a suspension of amine a (4.2 g, 20.9 mmol) in 6 N HCl (100 mL),cooled in ice-water bath, was added NaNO₂ (2.2 g, 31.4 mmol) in portions(caution was used due to elevated reaction temperature). After stirredat ice-water temperature for 1 h, this cold solution was added drop wiseonto an ice-cold solution of CuBr (5.4 g, 37.6 mmol) in water (150 mL).After addition, CH₂Cl₂ (80 mL) was added slowly to the mixture. Thereaction mixture was allowed to reach RT and was stirred for 4 h. It wasdiluted with more CH₂Cl₂ (50 mL). The phases were separated. The aqueousphase was extracted with CH₂Cl₂ (2×50 mL). The combined CH₂Cl₂ werewashed with sat. sodium thiosulfate (100 mL), dried (MgSO₄), andconcentrated. The crude product was adsorbed on to Celite and purifiedby ISCO CombiFlash 120 g column (1-8% ethyl acetate-hexane) to afford3.1 (66%) of chloro ester b as a white solid.

Example 17

Following the general procedure of Williams et al. (Tetrahedron Lett.1995, 36(31), 5461-5464), to a stirred suspension of ester a (1.5 g, 5.7mmol) and amine (700 mg, 7.1 mmol) in THF (30 mL) at −5° C. under N₂ wasadded CH₃MgCl (16.1 mL of 3 M solution in THF, 48.4 mmol) over 20 minwhile keeping the temperature below 0° C. After 0.5 h at −5° C., thereaction mixture was allowed to warm to RT and stirred for overnight.The reaction was quenched with 1 N HCl, diluted with 1 N HCl (100 mL),heated the mixture at 35° C. for 3 h, then cooled, diluted with EtOAc(150 mL), dried (MgSO₄), and concentrated in vacuo. The crude productwas adsorbed on to Celite and purified by ISCO CombiFlash 40 g column(1-10% ethyl acetate-hexane) to afford 900 mg (64%) of chloro ketone bas a clear liquid. Compound b may also be prepared employing theprocedures described by Alvaro et al. (WO 2004099143) and Tsuno et al.(Bull Chem. Soc. of Japan 1975, 48(11), 3347-55).

Example 18

Concentrated sulfuric acid (2 mL) was added slowly to a stirred solutionof 2-fluorophenylacetic acid a (10.0 g, 65.0 mmol) in toluene (100 mL)and EtOH (7.6 mL, 130 mmol). The resulting mixture was heated at 100° C.for 1.5 h. It was concentrated in vacuo, diluted with EtOAc (200 mL),washed with 10% K₂CO₃ until the washes were basic, dried (MgSO₄), andconcentrated in vacuo to afford 9.9 g (84%) of ester b as a light yellowoil.

Example 19

Concentrated sulfuric acid (3 mL) was added slowly to a stirred solutionof 2,4 di-fluorophenylacetic acid a (10.0 g, 58.1 mmol) in EtOH (100mL), stirred at RT for 2 d. It was concentrated, diluted with EtOAc (200mL), washed with 10% K₂CO₃ until the washes were basic, dried (MgSO₄),and concentrated in vacuo to afford 11.1 g (95.5%) of difluoro ester bas a white solid.

Example 20

Following the procedure for preparing ester from example 19,2-nitrophenylacetic acid a (10.0 g, 55.2 mmol) afforded 10.9 g (95%) ofnitro ester b as a light yellow solid.

Example 21

Following the general procedure described by Kemp et al. (J. Am. Chem.Soc. 1975, 97, 7305-7312), reaction of difluoro-ester a (4.0 g, 20.0mmol), isoamyl-nitrite (3.2 mL, 24.0 mmol), and NaOEt (1.4 g, 20.0 mmol)in EtOH (40 mL), after purified by ISCO CombiFlash 80 g column (2-30%ethyl acetate-hexane) to afford 2.1 g (47%) of difluoro oxime b as alight yellow solid.

Example 22

A solution of the nitro oxime a (5.0 g, 21.0 mmol) prepared according tothe procedures described by Kemp et al. (J. Am. Chem. Soc. 1975, 97,7305-7312) in DMF (30 mL) was added drop wise over 25 min to avigorously stirred suspension of hexane-washed NaH (60% in mineral oil,840 mg, 21.0 mmol) in DMF (40 mL) under N₂. The resulting dark coloredsolution was heated slowly to 130° C. for 8 h. It was diluted with water(200 mL), extracted with EtOAc (2×200 mL), washed the EtOAc with brine,dried (MgSO₄), and concentrated in vacuo. The crude product was adsorbedon to Celite and purified by ISCO CombiFlash 120 g column (1-10% ethylacetate-hexane) to afford 1.6 g (41%) of benzisoxazole b as an off whitesolid.

Example 23

A mixture of benzisoxazole acid a (1.23 g, 7.5 mmol) prepared accordingto the procedures described by Kemp et al. (J. Am. Chem. Soc. 1975, 97,7305-7312), amine HCl salt (736 mg, 7.5 mmol), EDC (1.44 g, 7.5 mmol),and DIPEA (1.2 mL, 6.7 mmol) in MeCN (50 mL) was stirred at RT forovernight. It was concentrated in vacuo, dissolved in EtOAc (200 mL),washed with 0.5 N HCl and water, dried (MgSO₄), and concentrated toafford 1.4 g (88%) of benzisoxazole amide b as an off white solid.

Example 24

Following the procedure for preparing bromo ketone from example 12, tobenzisoxazole amide a (1.2 g, 5.9 mmol) was added CH₃MgCl (6.0 mL of 3 Msolution in THF, 17.8 mmol). The crude product was adsorbed on to Celiteand purified by ISCO CombiFlash 40 g column (1-5% ethyl acetate-hexane)to afford 670 mg (71%) of benzisoxazole ketone b as a white crystalline.Compound b may also be prepared according to the procedures described bySmalley et al. (Science of Synthesis 2002, 11, 289-335) and Farooq etal. (WO 9614305).

Example 25

Bromine (184 μL, 3.6 mmol) was added drop wise to a solution ofbenzisoxazole ketone a (525 mg, 3.3 mmol) in AcOH (1.5 mL) and CH₂Cl₂(6.0 mL). After 1 h at RT, LCMS indicated no reaction. Five drops ofconc. HCl were added to the reaction mixture and stirred at RT forovernight. It was quenched with 10% Na₂S₂O₃, diluted with CH₂Cl₂ (100mL), washed with 5% NaHCO₃, separated, dried (MgSO₄), and concentratedin vacuo to afford 820 mg (105%) of bromo ketone b as a brown oil.

Example 26

Following the general procedure of Strupczewski et al. (J. Med. Chem.1985, 28, 761-769), to a suspension of NaH (60% in mineral oil, 37 mg,0.92 mmol) in THF (3.0 mL) was added drop wise a solution of difluorooxime a (140 mg, 0.61 mmol) in DMF (1.5 mL). The resulting mixture washeated at 70° C. for 4 h. It was cooled, poured onto water (30 mL),extracted with EtOAc (2×50 mL). The EtOAc was washed with water, dried(MgSO₄), and concentrated. The crude product was adsorbed on to Celiteand purified by ISCO CombiFlash 12 g column (1-5% ethyl acetate-hexane)to afford 60 mg (47%) of benzisoxazole ester b as an off white solid.

Example 27

A suspension of benzisoxazole a (1.6 g, 7.8 mmol) in 70% H₂SO₄ (30 mL)was heated at 80° C. for 4 h. It was cooled, poured onto crushed ice.The solid was collected by filtration, washed with water, and dried toafford 1.3 g (89%) of benzisoxazole acid b as a white solid.

Example 28

Following the procedure for preparing amide from example 23,benzisoxazole acid a (1.3 g, 7.0 mmol) afforded, after purified by ISCOCombiFlash 12 g column (2-15% ethyl acetate-hexane), 740 mg (47%) ofbenzisoxazole amide b as a white solid.

Example 29

Following the procedure for preparing of ketone from example 24,benzisoxazole amide a (740 mg, 3.3 mmol) afforded 390 mg (66%) ofbenzisoxazole ketone b as an off white solid.

Example 30

Following the procedure for preparing ester from example 19,2,5-difluorophenylacetic acid a (9.56 g, 55.6 mmol) afforded 9.24 g(83%) of difluoro ester b as a clear liquid.

Example 31

Following the procedure for preparing ester from example 19,2,3-difluorophenylacetic acid a (10.0 g, 58.1 mmol) afforded 10.8 g(93%) of difluoro ester b as a clear liquid.

Example 32

Following the procedure for preparing oxime from example 21, difluoroester a (9.2 g, 46.0 mmol) afforded 5.57 g (53%) of difluoro oxime b asa white solid.

Example 33

Following the procedure for preparing oxime from example 21, difluoroester a (10.8 g, 54 mmol) afforded 4.9 g (40%) of difluoro oxime b as awhite solid.

Example 34

Following the procedure for preparing ester from example 26, difluorooxime a (5.5 g, 24.0 mmol) afforded, after purified by ISCO CombiFlash40 g column (1-5% ethyl acetate-hexane), 2.66 g (53%) of benzisoxazoleester b as an off white solid.

Example 35

Following the procedure for preparing ester from example 26, difluorooxime a (4.9 g, 21.4 mmol) afforded 2.9 g (65%) of benzisoxazole ester bas a light yellow crystalline.

Example 36

Following the procedure for preparing acid from example 27,benzisoxazole ester a (2.1 g, 10.0 mmol) afforded 1.92 g (86%) ofbenzisoxazole acid b as an off white solid.

Example 37

Following the procedure for preparing acid from example 27,benzisoxazole ester a (2.4 g, 11.5 mmol) afforded 1.92 g (76%) ofbenzisoxazole acid b as an off white solid.

Example 38

Following the procedure for preparing amide from example 23,benzisoxazole acid a (1.4 g, 7.73 mmol) afforded 1.95 g (83%) ofbenzisoxazole amide b as a yellow solid.

Example 39

(Following the procedure for preparing amide from example 23,benzisoxazole acid a (1.9 g, 10.5 mmol) afforded 1.7 g (72%) ofbenzisoxazole amide b as a brown solid.

Example 40

Following the procedure for preparing ketone from example 12,benzisoxazole amide a (1.95 g, 8.7 mmol) afforded 448 mg (30%) ofbenzisoxazole ketone b as a brown oil. Compound b may also be preparedaccording to the procedures described by Farooq et al. (WO 9614305).

Example 41

Following the procedure for preparing ketone from example 12,benzisoxazole amide a (1.70 g, 7.6 mmol) afforded 192 mg (14%) ofbenzisoxazole ketone b as a white crystalline solid.

Example 42

A suspension of 1,4-naphthalenedicarboxylic acid a (10.0 g, 46.3 mmol)in MeOH (70 mL) and H₂SO₄ (5 mL) was stirred at RT for 2 days, and thenheated at 50° C. for 10 h. It was concentrated in vacuo, re-dissolved inCH₂Cl₂ (300 mL), washed with 10% K₂CO₃ (200 mL), dried (MgSO₄), andconcentrated to give 6.6 g (60%) of di-ester b as yellow solid.

Example 43

A mixture of the di-ester a (2.0 g, 8.2 mmol), LiOH (344 mg, 8.2 mmol)in THF (40 mL), water (5 mL), and MeOH (1 mL) was stirred at RTovernight. LCMS indicated some starting di-ester still remainsun-reacted. Extra LiOH (84 mg, 2.0 mmol) was added to the reactionmixture. After 4 h, it was diluted with 0.5 N HCl (100 mL), extractedwith EtOAc (100 mL), dried (MgSO₄), concentrated to afford 1.4 g (74%)of monoacid b as a light yellow solid.

Example 44

A mixture of monoacid a (1.4 g, 6.1 mmol) and SOCl₂ (9 mL) in toluene(15 mL) was heated at 75° C. for 4 h. The solvent was removed in vacuo,diluted with toluene (50 mL) and concentrated, and dried under highvacuum overnight. The residue was suspended in toluene (30 mL) andcooled in ice-water bath. Me₂Zn (12 mL of 1 M solution in heptane, 12.0mmol) was added slowly, stirred at RT for 3.5 h. The reaction wasquenched with sat. NH₄Cl, diluted with water (100 mL), extracted withEtOAc (2×100 mL), dried (MgSO₄), and concentrated in vacuo. The crudeproduct was adsorbed on to Celite and purified by ISCO CombiFlash 40 gcolumn (1-10% ethyl acetate-hexane) to afford 890 mg (86%) of ketoneester b as an off white solid. Compound b may also be prepared accordingto the procedures described by Uehata et al. (JP 2003073357).

Example 45

Bromine (1.49 g, 9.3 mmol) was added slowly to a stirred solution ofketone a (1.47 g, 8.5 mmol) prepared according to the proceduresdescribed by Berg et al. (WO 02066480) in 33% HBr/AcOH (20 mL) at RT. Itwas stirred for 1 h, diluted with ether (65 mL), and stirred vigorouslyfor 1 h. The solid was collected by filtration, washed with ether, highvacuum dried to afford 2.88 g (100%) of bromo methyl ketone b as ayellow solid.

Example 46

A mixture of 2-hydroxyquinoline-4-carboxylic acid a (6.25 g, 33.1 mmol),MeI (10.33 g, 72.7 mmol), and K₂CO₃ (10.0 g, 72.7 mmol) in DMF (110 mL)was heated at 80° C. for 16 h overnight. LCMS indicated incompletereaction. Extra MeI (4.69 g, 33.1 mmol) was added to the reactionmixture and heated at 100° C. for 3 h. It was cooled, poured into icewater, and 10% K₂CO₃ (50 mL), extracted with EtOAc (2×150 mL). Thecombined EtOAc were washed with water, dried (MgSO₄), concentrated invacuo. The crude product was adsorbed on to Celite and purified by ISCOCombiFlash 80 g column (2-50% ethyl acetate-hexane) to afford 4.68 g(65%) of dihydroquinoline ester b as an off white solid.

Example 47

Lithium hydroxide (1.45 g, 34.5 mmol) was added to a solution ofdihydroquinoline ester a (1.50 g, 6.91 mmol) in THF (40 mL), followed bywater (10 mL). The mixture was stirred at RT for 16 h. It wasconcentrated in vacuo, diluted with EtOAc (100 mL) and 0.5 N HCl (100mL). A white solid precipitated and was collected by filtration, washedwith water. The EtOAc was separated, dried (MgSO₄), concentrated invacuo to afford a white solid product. A combined yields of 1.41 g(100%) of dihydroquinoline acid b as a white solid, which was used without further purification.

Example 48

N,N-Diisopropylethylamine (1.37 g, 10.7 mmol) was added to a suspensionof dihydroquinoline acid a (2.42 g, 11.9 mmol), amine (1.40 g, 14.3mmol), and EDC (2.28 g, 11.9 mmol) in MeCN (80 mL), stirred at RT for 2h. It was concentrated in vacuo, diluted with CH₂Cl₂ (100 mL), washedwith 0.5 N HCl (50 mL) and 0.5 N NaOH (50 mL), dried (MgSO₄),concentrated to afford 1.98 g (67%) of dihydroquinoline amide b as awhite solid, which was used with out further purification.

Example 49

Following the procedure for preparing ketone from example 12,dihydroquinoline amide a (2.17 g, 8.82 mmol), CH₃MgCl (8.82 mL of 3 Msolution in THF, 26.5 mmol) afforded 766 mg (43%) of dihydroquinolineketone b as a yellow solid. Compound b may also be prepared according tothe procedures described by Fujita et al. (Chem. & Pharm. Bull 2001,49(7), 900-904 and Chem. & Pharm. Bull 2001, 49(4), 407-412).

Example 50

Following the general procedure of Legros et al. (Tetrahedron 2001, 57,2507-2514), 4-bromoisoquinoline a (1.0 g, 4.8 mmol) afforded 707 mg(86%) of ketone b as a light yellow solid.

Example 51

Following the procedure for preparing amide from example 48,dihydroisoquinoline acid a (1.25 g, 6.2 mmol) prepared according to theprocedures described by Deady et al. (J. Heterocyclic Chem. 2001, 38,1185) afforded 754 mg (50%) of dihydroisoquinoline amide b as a yellowgum.

Example 52

Following the procedure for preparing ketone from example 49,dihydroisoquinoline aimde a (0.754 g, 3.06 mmol) afforded 510 mg (85%)of dihydroisoquinoline ketone b as a yellow solid. Compound b may alsobe prepared according to the procedures described by Alvarez et al.(Science of Synthesis 2005, 15, 839-906), Kimura et al. (Chem. & Pharm.Bull 1983, 31(4), 1277-82), Tomisawa et al. (Chem. & Pharm. Bull 1975,23(3), 592-6) and Dyke et al. (Tetrahedron 1973, 29(23), 3881-8).

Example 53

A mixture of 2-hydroxyquinoline-4-carboxylic acid a (4.0 g, 21.2 mmol),POBr₃ (25.0 g, 87.2 mmol) in toluene (40 mL) was heated at 100° C. for 3h. It was cooled to RT, carefully poured onto crushed ice, extractedwith EtOAc (2×250 mL), dried (MgSO₄), concentrated in vacuo. The residuewas dissolved in 1 N NaOH (150 mL), extracted with EtOAc (2×100 mL). Theaqueous layer was then acidified with 1 N HCl to pH 3. The white solidwas collected by filtration, washed with water, dried to afford 3.0 g(56%) of bromo acid b as a white solid.

Example 54

Following the procedure for preparing amide from example 48, bromo acida (3.0 g, 11.9 mmol) afforded 2.67 g (77%) of bromo amide b as a whitesolid.

Example 55

(46801-78) Following the procedure for preparing ketone from example 49,bromo amide a (1.0 g, 3.4 mmol) afforded 800 mg (94%) of bromo ketone bas an off white solid.

Example 56

Trifluoromethane sulfonic anhydride (6.82 g, 24.2 mmol) was added dropwise onto to a mixture of ethyl-4-hydroxyquinoline carboxylate a (5.0 g,23.0 mmol) and pyridine (1.95 mL, 24.2 mmol) in CH₂Cl₂ (100 mL) at icewater bath temperature under N₂. The mixture was stirred at RTovernight. It was diluted with CH₂Cl₂ (100 mL), washed with 0.5 N NaOH(100 mL), dried (MgSO₄), concentrated in vacuo. The crude product wasadsorbed on to Celite and purified by ISCO CombiFlash 80 g column (2-15%ethyl acetate-hexane) to afford 7.4 g (93%) of triflate b as a whitesolid.

Example 57

Following the general procedure of Legros et al. (Tetrahedron 2001, 57,2507-2514), a mixture of triflate a (1.0 g, 2.86 mmol),bis(dibenzylideneacetone)palladium(0) (82 mg, 0.14 mmol),1,3-bis(diphenylphosphino)propane (65 mg, 0.16 mmol), and Et₃N (1.19 mL,8.58 mmol) in DMF (10 mL) were stirred at RT for 15 min under N₂.n-Butyl vinyl ether (1.43 g, 14.3 mmol) in DMF (5 mL) was added and theresulting mixture was stirred at 80° C. for 24 h. It was cooled to RT, 1N HCl (30 mL) were added slowly, stirred at RT for 24 h. The mixture wasneutralized with 1 N NaOH and extracted with ether (2×100 mL), dried(MgSO₄), and concentrated. The crude product was adsorbed on to Celiteand purified by ISCO CombiFlash 12 g column (2-20% ethyl acetate-hexane)to afford 460 mg (66%) of ketone b as a white solid.

Example 58

A mixture of ketone ester a (1.50 g, 6.17 mmol), KOH (640 mg, 11.35mmol) in EtOH (30 mL) was stirred at RT overnight. The reaction mixturewas diluted with water (150 mL), extracted with EtOAc (100 mL). Theaqueous layer was then acidified with 1 N HCl, extracted with EtOAc(2×100 mL), dried (MgSO₄), concentrated in vacuo afforded 1.53 g (100%)of ketone acid b as a yellow solid. Compound b may also be preparedaccording to the procedures described by Priestly et al. (Bioorg. & Med.Chem. 1996, 4(7), 1135-1147).

Example 59

A mixture of the ketone acid a (1.30 g, 5.16 mmol), dimethylaminehydrochloride (480 mg, 5.93 mmol), EDC (1.14 g, 5.93 mmol), and DIPEA(765 mg, 5.93 mmol) in MeCN (30 mL) was stirred at RT overnight. Solventwas removed. The residue was dissolved in CH₂Cl₂ (100 mL), washed with 1N HCl (50 mL) and 1 N NaOH (50 mL), dried (MgSO₄), and concentrated. Thecrude product was adsorbed on to Celite and purified by ISCO CombiFlash40 g column (2-20% ethyl acetate-dichloromethane) to afford 656 mg (53%)of ketone amide b as a light yellow gum.

Example 60

Following the procedure for preparing triflate from example 56,5-hydroxyquinoline a (3.42 g, 23.6 mmol) afforded 6.0 g (92%) oftriflate b as light yellow liquid.

Example 61

Following the procedure for preparing ketone from example 57, triflate a(6.0 g, 21.7 mmol) afforded 3.58 g (97%) of ketone b as a brown oil.

Example 62

Following the procedure for preparing triflate from example 56,2-(trifluoromethyl)-4-hydroxyquinoline a (6.87 g, 32.3 mmol) afforded9.31 g (84%) of triflate b as a yellow solid.

Example 63

Following the procedure for preparing ketone from example 57, triflate a(7.35 g, 21.3 mmol) afforded 1.79 g (35%) of ketone b as a yellow solid.

Example 64

Following the procedure for preparing amide from example 59,2-phenyl-4-quinolinecarboxylic acid a (5.0 g, 20.1 mmol) afforded 3.29 g(56%) of amide b as an off white solid.

Example 65

Following the procedure for preparing ketone from example 49, amide a(3.29 g, 11.26 mmol) afforded 3.29 g (118%) of ketone b as a yellowsolid. Compound b may also be prepared according to the proceduresdescribed by Sato et al. (JP 2002371078), Wong et al (WO 9846572),Leardini et al. (J. Chem. Soc., Chem. Communications 1984, 20, 1320-1),Kaneko et al. (Chem. & Pharm. Bull 1982, 30(1), 74-85), Schwenk et al.(J. Org. Chem. 1946, 11, 798-802) and Shivers et al (J. Am. Chem. Soc.1947, 69, 119-23).

Example 66

Following the procedure for preparing amide from example 59,2-hydroxy-4-quinolinecarboxylic acid a (5.0 g, 26.4 mmol) afforded 1.88g (30%) of amide b as a cream color solid.

Example 67

Following the procedure for preparing ketone from example 49, amide a(1.88 g, 8.1 mmol) afforded 993 mg (66%) of ketone b as a light yellowsolid. Compound b may also be prepared according to the proceduresdescribed by Wetzel et al. (J. Med. Chem. 1973, 16(5), 528-32), Jones etal. (J. Chem. Soc. [Section C]: Organic 1967, 19, 1808-13) and Ochia etal. (Chem. & Pharm. Bull 1963, 11, 137-8).

Example 68

Boc-ester a (900 mg, 2.0 mmol) was dissolved in THF (20 mL) and MeOH (1mL) at ice water bath temperature. NaBH₄ (300 mg, 8.0 mmol) was addedand the mixture was stirred for 1 h then another 1 h at RT. Reaction wasquenched with the addition of few drops of water, then diluted with morewater (100 mL), extracted with EtOAc (2×100 mL), dried (MgSO₄), andconcentrated in vacuo to afford 739 mg (90%) of alcohol b as a yellowfoamy solid.

Example 69

To the mixture of amine a (159 mg, 0.46 mmol), Boc-cyclohexyl-Gly-OH b(129 mg, 0.50 mmol), and HATU (350 mg, 0.92 mmol) in MeCN (4 mL) wasadded DIPEA (162 μL. The resulting mixture was stirred at RT for 2 h.LCMS indicated incomplete reaction. An extra equivalence of HATU (175mg, 0.46 mmol) and DIPEA (81 μL, 0.46 mmol) were added and stirred foranother 1 h. Solvent was removed in vacuo, diluted with CH₂Cl₂ (10 mL),washed with 0.5 N HCl (10 mL) and with water, dried (MgSO₄), andconcentrated. The crude product was adsorbed on to Celite and purifiedby ISCO CombiFlash 12 g column (0-5% MeOH/CH₂Cl₂) to afford 187 mg (74%)of product c as a brown solid.

Example 70

To the mixture of amine a (62 mg, 0.13 mmol), Boc-N-Me-Ala-OH b (27 mg,0.13 mmol), and HATU (99 mg, 0.26 mmol) in MeCN (2 mL) was added DIPEA(46 μL, 0.26 mmol). The resulting mixture was stirred at RT for 2 h.Solvent was removed in vacuo, diluted with CH₂Cl₂ (10 mL), washed with0.5 N HCl (10 mL) and with water, dried (MgSO₄), and concentrated toafford 69 mg (85%) of product c as a clear oil.

Example 71

A mixture of di-peptide a (100 mg, 0.29 mmol), thiazole amine b (113 mg,0.32 mmol), HOAt (59 mg, 0.435 mmol), and DIC (67 μL, 0.435 mmol) inCH₂Cl₂ (3 mL) was stirred at RT for overnight. The mixture was dilutedwith CH₂Cl₂ (10 mL), washed with 0.5 N HCl (10 mL) and 0.5 N NaOH (10mL), dried (MgSO₄), and concentrated. The crude product was adsorbed onto Celite and purified by ISCO CombiFlash 12 g column (10-90% ethylacetate-hexane) to afford 175 mg (89%) of product c as a clear oil.

Example 72

Boc-amine a (175 mg, 0.26 mmol) was treated with (1:1) TFA/CH₂Cl₂ (8mL), catalytic toluene at RT for 1 h. Solvent was removed in vacuo. Theresidue was purified by reverse-phase HPLC (Cl₈, MeCN—H₂O, 0.1% TFA) andlyophilized to afford 98 mg (49%, in 2 steps) of desired product b as ahygroscopic white solid.

Example 73

A mixture of the Boc-ester a (200 mg, 0.30 mmol), LiOH (200 mg, 0.30mmol) in THF (2 mL), and water (25 μL) was stirred at RT for 1 h. MeOH(500 μL) was added and stirred at RT overnight. It was diluted withEtOAc (10 mL), washed with 0.5 N HCl (10 mL), dried (MgSO₄),concentrated in vacuo to afford 160 mg (82%) of Boc acid 187 as a whitesolid.

Example 74

A mixture of the Boc nitrile a (200 mg, 0.31 mmol), NaN₃ (309 mg, 4.75mmol), and NH₄Cl (252 mg, 4.75 mmol) in DMF (3 mL) was heated at 100° C.for 3.5 d. It was cooled to RT, diluted with water, extracted with EtOAc(2×50 mL), washed with brine, dried (MgSO₄), concentrated in vacuo. Thecrude product was adsorbed on to Celite and purified by ISCO CombiFlash4 g column (10-90% ethyl acetate-hexane) to afford 37 mg of tetrazole #as a brown oil. More material was recovered by extracting the aqueouslayer with CH₂Cl₂ to recover 65 mg of product. A combined 102 mg (49%)of tetrazole b was isolated.

Example 75

A mixture of the Boc ester a (135 mg, 0.20 mmol), KOH (14.5 mg, 0.26mmol) in EtOH (4 mL) was stirred at RT for 2 h, then diluted with EtOAc(8 mL), acidified with 1 N HCl, separated, dried (MgSO₄), concentratedin vacuo, and high vacuum dried. The crude product b was carried onwithout further purification.

Example 76

Following the general procedure of Yamamoto [Asao, N; Lee, S.; Yamamoto,Y. Tetrahedron Letters, 2003, 4265-4266.], MeLi (20 mL of 1.6M solutionin ether, 30.2 mmol) was added to a 0° C. suspension of powdered CuI(2.90 g, 15.1 mmol) and ether (5 mL). The resulting grey solution wasstirred vigorously for 10 min then concentrated under reduced pressureat 0° C. Dichloromethane (20 mL, precooled to 0° C.) was added, then thesuspension was cooled to −78° C. and TMSCl (1.9 mL, 15.1 mmol) was addedfollowed quickly by a solution of ester 192 [WO 01168603] (1.0 g, 5.0mmol) and CH₂Cl₂ (50 mL), the mixture was stirred vigorously at −78° C.for 30 min, then at 0° C. for 2 h. The mixture was then poured into 200mL of 1:1 saturated NH₄Cl:NH₄OH. The layers were separated and theaqueous phase was extracted with CH₂Cl₂ (2×20 mL). The combined organicphases were washed with brine (1×20 mL), dried (Na₂SO₄), adsorbed ontoCelite and purified by chromatography ISCO CombiFlash 12 g column, 0-8%ethyl acetate-hexanes to afford 783 mg (72%) of ester 193 as a clearoil.

Example 77

A solution of ester a (780 mg), CH₂Cl₂ (5 mL), and TFA (2 mL) wasmaintained at rt overnight. The solvents were removed under reducedpressure to afford quantitative yield acid b as a colorless oil whichwas used without further purification.

Example 78

Following the general procedure of Evans [Evans, D. A.; Britton, T. C.;Ellman, J. A.; Dorow, R. L. J. Am. Chem. Soc. 1990, 112, 4011-4030],pivaloyl chloride (3.2 mL, 26 mmol) was added to a −10° C. solution ofacid a (3.72 g, 23.5 mmol), TEA (4.3 mL, 31 mmol) and THF (50 mL). Theresulting white slurry was allowed to warm to −5° C. over 20 min withvigorous stirring. The mixture was then cooled to −78° C. and a solutionof the lithium salt of (S)-4-benzyl-2-oxazolidinone (prepared from(S)-4-benzyl-2-oxazolidinone (7.5 g, 42.3 mmol), n-BuLi (26 mL of 1.6Msolution in hexanes, 42.3 mmol) and THF (150 mL) at −78° C.) was addedvia cannula over 10 min. The mixture was maintained at −78° C. for 1 h,then quenched with saturated NH₄Cl (200 mL) and the THF removed underreduced pressure. The aqueous phase was extraced with EtOAc (3×50 mL).The combined organic phases were washed with brine (1×50 mL), dried(Na₂SO₄), adsorbed onto Celite and purified by chromatography ISCOCombiFlash 120 g column, 5-40% ethyl acetate-hexanes to afford 5.7 g(76%) of imide b as a clear oil.

Example 79

Following the general procedure of Evans [Evans, D. A.; Britton, T. C.;Ellman, J. A.; Dorow, R. L. J. Am. Chem. Soc. 1990, 112, 4011-4030], acold (−78° C.) solution of imide a (5.7 g, 18 mmol) and THF (64 mL) wasadded to a cold (−78° C.) solution of KHMDS (20 mmol) and THF (120 mL)over 10 min. The colorless solution was maintained at −78° C. for 30min., then a cold (−78° C.) solution of TrsylN₃ (6.9 g, 22.4 mmol) andTHF (40 mL) was added via cannula over 5 min. Acetic acid (5.3 mL, 90mmol) was added and the mixture was immediately brought to 30° C. andheld there for 1 h. The reaction was quenched with brine (200 mL) andCH₂Cl₂ (200 mL). The phases were separated and the aqueous phase wasextraced with CH₂Cl₂ (2×50 mL). The combined organic phases were washedwith saturated NaHCO₃ (1×50 mL), dried (Na₂SO₄), adsorbed onto Celiteand purified by chromatography ISCO CombiFlash 330 g column, 5-45% ethylacetate-hexanes to afford 4.2 g (65%) of azo imide b as a colorlesssolid.

Example 80

A mixture of azido-imide a (4.7 g, 13 mmol) LiOH.H₂O (660 mg, 15.6 mmol)THF (93 mL) and water (31 mL) was maintained at rt for 1 d. FurtherLiOH.H₂O (200 mg) was added, and the mixture stirred for 2 h. SolidNaHCO₃ (2.18 g) was added and the THF removed under reduced pressure.Following dilution with water (150 mL), the aqueous phase was washedwith CH₂Cl₂ (3×50 mL) and the combined organic phases were extractedwith saturated NaHCO₃ (1×50 mL). The combined aqueous phases wereacidified with con. HCl to pH<2 and extracted with EtOAc (4×50 mL). Thecombined organic phases were dried (Na₂SO₄), and concentrated to yield1.38 g (53%) of acid b as a colorless solid.

Example 81

Isoquinoline carboxylic acid a (5.0 g, 28.9 mmol),N,O-dimethy-hydroxylamine hydrochloride (3.1 g, 31.8 mmol) EDC (6.1 g,32 mmol), DIPEA (5.7 mL, 32 mmol) and MeCN (50 mL) were mixed togetherand stirred at rt overnight. The MeCN was removed under reduced pressureand the residue partitioned between water (200 mL) and EtOAc (200 mL).The phases were separated and the aqueous phase was extracted with EtOAc(2×50 mL). The combined organic phases were b as a colorless solid.

Example 82

Methyl magnesium chloride (12.3 mL of 3.0M THF) was added to a 0° C.solution of amide a (4.0 g, 18.5 mmol) and THF (40 mL). After 30 min at0° C., the cooling bath was removed for 40 min. The reaction was pouredinto cold saturated NH₄Cl (200 mL), and extracted with EtOAc (3×50 mL).The combined organic phases were washed with water, brine, dried(Na₂SO₄), and concentrated to yield 3.15 g (100%) of ketone b as acolorless oil.

Example 83

Following the general procedure of Barlin [Barlin, G. A.; Davies, L. P.;Ireland, S. J.; Ngu, M. M. L. Aust. J. Chem. 1989, 42, 1735-1748], Br₂(150 μL, 2.92 mmol) was added in one portion to a solution of ketone a(500 mg, 2.92 mmol) and 33% HBr/AcOH (10 mL). After 1 h, ether (20 mL)was added, and the ppt was collected on filter paper, washed with ether,and dried under vacuum to afford 910 mg (94%) of bromide b as a yellowsolid.

Example 84

A mixture of 4-carboxy-2-hydroxyquinoline a (500 mg, 2.64 mmol) andPOCl₃ (5 mL) was heated at 100° C. for 1 h. The solvent was removedunder reduced pressure, and the residue was dissolved in CH₂Cl₂ (10 mL),and cooled to 0° C. Morpholine (1.0 mL, 13.2 mmol) was added dropwise,and the mixture was allowed to come to rt. The mixture was thenre-cooled to 0° C. and more morpholine (1.0 mL, 13.2 mmol) was addeddropwise, and the mixture was allowed to come to rt overnight. Themixture was then diluted with CH₂Cl₂ (50 mL), and washed with saturatedNH₄Cl (3×20 mL). The combined aqueous phases were extracted with CH₂Cl₂(1×20 mL), and the combined organic phases were dried (Na₂SO₄), adsorbedonto Celite and purified by chromatography ISCO CombiFlash 12 g column,5-75% ethyl acetate-hexanes to afford 570 mg (78%) of amide b as acolorless solid.

Example 85

Diethyl zinc (2.3 mL of 1.1M solution in toluene, 2.5 mmol) was added toa mixture of amide a (500 mg, 1.8 mmol), NiCl₂DPPP (100 mg, 0.18 mmol),and THF (5 mL) (caution was used due to exothermic reaction). The darksolution was then heated at 100° C. in a μW reactor for 15 min. Thereaction was then quenched into saturated NH₄Cl (50 mL), and extractedwith EtOAc (3×20 mL). The combined organic phases were dried (Na₂SO₄),adsorbed onto Celite and purified by chromatography ISCO CombiFlash 12 gcolumn, 0-75% ethyl acetate-hexanes to afford 350 mg (71%) of amide b asa colorless solid.

Example 86

Diisopropyl zinc (3 ml of 10M solution in toluene, 2.5 mmol) was addedto a mixture of chloride a (500 mg, 1.8 mmol), NiCl₂DPPP (115 mg, 0.18mmol), and THF (3 mL) The dark solution was then heated at 100° C. in aμW reactor for 15 min. The reaction was then quenched into saturatedNH₄Cl (50 mL), and extracted with EtOAc (3×20 mL). The combined organicphases were dried (Na₂SO₄), adsorbed onto Celite and purified bychromatography ISCO CombiFlash 12 g column, 0-15% ethyl acetate-hexanesto afford 383 mg (74%) of amide b as a colorless solid.

Example 87

Methyl magnesium chloride (12.3 mL of 3.0M in THF, 37 mmol) was added to0° C. solution of amide a (2.84 g, 10.51 mmol) and THF (20 mL). Thesolution was allowed to come to rt, then maintained at that temp. for 4h. The reaction was quenched into cold saturated NH₄Cl (100 mL),extracted with EtOAc (3×50 mL). The combined organic phases were washedwith brine (1×50 mL), dried (Na₂SO₄), adsorbed onto Celite and purifiedby chromatography ISCO CombiFlash 40 g column, 0-30% ethylacetate-hexanes to afford 1.88 g (86%) of ketone b as a colorless oil.

Example 88

Following the general procedure of Barlin [Barlin, G. A.; Davies, L. P.;Ireland, S. J.; Ngu, M. M. L. Aust. J. Chem. 1989, 42, 1735-1748], Br₂(640 μL, 2.92 mmol) was added in one portion to a solution of ketone a(2.27 g, 11.4 mmol) and 33% HBr/AcOH (40 mL). After 1 h, ether (50 mL)was added, and the ppt was collected on filter paper, washed with ether,and dried under vacuum to afford 3.88 g (94%) of bromide b as a yellowsolid.

Example 89

Following the general procedure of Rieke [Zhu, L.; Wehmeyer, R. M.;Rieke, R. D. J. Org. Chem. 1991, 56, 1445-1453], propyl zinc bromide(4.0 mL of 0.5M solution in THF, 2.0 mmol) was added to a mixture ofchloride a (500 mg, 1.81 mmol), Pd(PPh₃)₄ (100 mg, 0.09 mmol) and THF (3mL). The resulting solution was heated at 70° C. in a μW reactor for 15min. The reaction was then quenched into saturated NH₄Cl (50 mL), andextracted with EtOAc (3×20 mL). The combined organic phases were dried(Na₂SO₄), adsorbed onto Celite and purified by chromatography ISCOCombiFlash 12 g column, 0-75% ethyl acetate-hexanes to afford 400 mg(77%) of amide b as a colorless solid.

Example 90

Following the general procedure of Rieke [Zhu, L.; Wehmeyer, R. M.;Rieke, R. D. J. Org. Chem. 1991, 56, 1445-1453], cyclopently zincbromide (4.0 mL of 0.5M solution in THF, 2.0 mmol) was added to amixture of chloride a (500 mg, 1.81 mmol), Pd(PPh₃)₄ (100 mg, 0.09 mmol)and THF (3 mL). The resulting solution was heated at 70° C. in a μWreactor for 15 min. The reaction was then quenched into saturated NH₄Cl(50 mL), and extracted with EtOAc (3×20 mL). The combined organic phaseswere dried (Na₂SO₄), adsorbed onto Celite and purified by chromatographyISCO CombiFlash 12 g column, 0-75% ethyl acetate-hexanes to afford 333mg (59%) of amide b as a colorless solid.

Example 91

Methyl magnesium chloride (7.3 mL of 3.0M in THF, 22 mmol) was added to0° C. solution of amide a (1.77 g, 6.2 mmol) and THF (15 mL). Thesolution was allowed to come to rt, then maintained at that temp. for 4h. The reaction was quenched into cold saturated NH₄Cl (100 mL),extracted with EtOAc (3×50 mL). The combined organic phases were washedwith brine (1×50 mL), dried (Na₂SO₄), adsorbed onto Celite and purifiedby chromatography ISCO CombiFlash 40 g column, 0-30% ethylacetate-hexanes to afford 1.14 g (85%) of ketone b as a colorless oil.

Example 92

Following the general procedure of Angibaud [Angibaud, P; et. al,Bioorg. Med. Chem. Lett. 2003, 13, 4365-4369], a mixture of chloride a(1.0 g, 5.0 mmol) NaN₃ (1.6 g, 25 mmol) DMF (10 mL) and water (1.0 mL)was heated at heated at 120° C. in a μW reactor for 2 h. The reactionwas then quenched into water (50 mL), and extracted with EtOAc (3×20mL). The combined organic phases were dried (Na₂SO₄), adsorbed ontoCelite and purified by chromatography ISCO CombiFlash 40 g column, 0-50%ethyl acetate-hexanes to afford 300 mg (28%) of tetrazole b as a yellowsolid.

Example 93

Trifluoromethane sulfonic anhydride (5.0 g, 17.7 mmol) was added dropwise to a mixture of 2-methyl-4-hydroxyquinoline a (2.56 g, 16.1 mmol)and pyridine (1.54 mL, 17.7 mmol) in CH₂Cl₂ (25 mL) at ice water bathtemperature under N₂. The mixture was allowed to warm to 10° C. It wasdiluted with CH₂Cl₂ (100 mL), washed with saturated NaHCO₃ (3×50 mL),dried (Na₂SO₄), adsorbed on to Celite and purified by ISCO CombiFlash 40g column (0-30% ethyl acetate-hexane) to afford 2.57 g (54%) of triflateb as a dark oil.

Example 94

Following the general triflation procedure, 7-chloro-4-hydroxyquinolinea (10.0 g, 35.4 mmol) afforded 7.5 g (68%) of triflate b as a colorlesssolid.

Example 95

Following the general triflation procedure 6-flouro-4-hydroxyquinoline a(5.0 g, 28.2 mmol) afforded 6.6 g (75%) of triflate b as a colorlesssolid.

Example 96

Following the general procedure of Legros [Tetrahedron 2001, 57,2507-2514], a mixture of triflate a (7.5 g, 24.1 mmol),bis(dibenzylideneacetone)palladium(0) (690 mg, 1.2 mmol),1,3-bis(diphenylphosphino)propane (546 mg, 1.33 mmol), and Et₃N (10 mL,72.3 mmol) in DMF (50 mL) were stirred at RT for 15 min under N₂.n-Butyl vinyl ether (15 mL, 120 mmol) in DMF (15 mL) was added and theresulting mixture was stirred at 80° C. for 24 h. It was cooled to RT, 1N HCl (150 mL) were added slowly, stirred at RT for 24 h. The mixturewas neutralized with 1 N NaOH and extracted with ether (3×100 mL), dried(MgSO₄), and concentrated. The crude product was adsorbed on to Celiteand purified by ISCO CombiFlash 120 g column (5-30% ethylacetate-hexane) to afford 1.62 g (32%) of ketone b as a white solid.

Example 97

Following the general procedure for preparing ketone from example 96,6.56 g of triflate a afforded 3.14 g (73%) of ketone b as a colorlesssolid.

Example 98

Following the general procedure for preparing ketone from example 96,2.57 g of triflate a afforded 820 mg (50%) of ketone b as a colorlesssolid.

Example 99

Methyl magnesium chloride (0.5 mL of 3.0M in THF, 1.5 mmol) was added to0° C. solution of ester a (230 mg, 0.5 mmol) and THF (5 mL). Thesolution was maintained at 0° C. for 2 h. The reaction was quenched intocold saturated NH₄Cl (50 mL), extracted with EtOAc (3×20 mL). Thecombined organic phases were washed with brine (1×50 mL), dried(Na₂SO₄), adsorbed onto Celite and purified by chromatography ISCOCombiFlash 12 g column, 0-50% ethyl acetate-hexanes to afford 135 mg(61%) of alcohol b as a colorless oil

Example 100

Bromine (122 μL, 2.4 mmol) was added to the solution of benzisoxazoleketone a (390 mg, 2.2 mmol) in AcOH (1.5 mL) and CH₂Cl₂ (6 mL). After 1h at RT, LCMS indicated no reaction. Four drops of conc. HCl were addedto the reaction mixture and stirred at RT overnight. It was quenchedwith 10% Na₂S₂O₃, diluted with CH₂Cl₂ (100 mL) and water, separated,washed the organic layer with 5% NaHCO₃, dried (MgSO₄), and concentratedto afford 537 mg (95%) of bromo ketone b as an off white solid.

Example 101

A mixture of bromo ketone a (537 mg, 2.1 mmol), thioamide b (718 mg, 3.1mmol), and pyridine (153 μL, 1.9 mol) in EtOH (15 mL) was heated at 70°C. for 1 h. It was concentrated in vacuo. The crude product was adsorbedon to Celite and purified by ISCO CombiFlash 40 g column (3-30% ethylacetate-hexane) to afford 190 mg (23%) of thiazole c as a light yellowgum.

Example 102 Tetrahydropyranylglycine

Tetrahydropyranylglycine was purchased from NovaBiochem, or synthesizedaccording to the literature: Ghosh, A. K.; Thompson, W. J.; holloway, M.K.; McKee, S. P.; Duong, T. T.; Lee, H. Y.; Munson, P. M.; Smith, A. M.;Wai, J. M; Darke, P. L.; Zugay, J. A.; Emini, E. A.; Schleife, W. A.;Huff, J. R.; Anderson, P. S. J. Med. Chem, 1993, 36, 2300-2310.

Example 103 Piperidinylglycine

Piperidinylglycine was synthesized according to the procedures describedby Shieh et al. (Tetrahedron: Asymmetry, 2001, 12, 2421-2425.

Example 104 4,4-Difluorocyclohexylglycine

4,4-difluorocyclohexylglycine was made according to the proceduresdescribed in US 2003/0216325.

Example 105 Boc (S)-2-amino-2-(4-hydroxycyclohexyl)acetic acid

Following the procedure described by Sheih et al. (Tetrahedron:Asymmetry, 2001, 12, 2421-2425), a solution of ketone a (8.4 g) andEtOAc (30 mL) was added to a solution of N-Cbz-phosphonoglycine methylester b, TMG (4.5 mL) and EtOAc (30 mL). The solution was maintained atrt for 48 h, then washed with 1N HCl (3×50 mL), brine (1×50 mL) dried(Na₂SO₄), filtered, and concentrated. The residue was adsorbed ontoCelite, and purified by chromatography, then further purified byre-crystalization from EtOAc/hexanes to afford 5.2 g of product c.

Following the procedure described by Sheih, (Tetrahedron: Asymmetry,2001, 12, 2421-2425), a solution of eneamide c (5.0 g),(S,S)-Me-BPE-Rh(I) (1.5 g, Strem Chemicals, Newburyport, Mass.), andMeOH (100 mL) was shaken vigorously under 70 psi of H₂ for 48 h. Thesolvent was removed under reduced pressure. The residue was taken up inEtOAc, and filtered through SiO₂ with more EtOAc. The solvent wasremoved under reduced pressure to afford 4.0 g of product d as acolorless solid.

A mixture of Cbz-carbamate d, (4.0 g) Boc₂O, (2.9 g), 20% Pd(OH)₂.C (1.0g) and MeOH (30 mL) was maintained under an atmosphear of H₂ for 6 h.The mixture was filtered through Celite with MeOH. The solvent wasremoved under reduced pressure to afford 4.5 g of residue e, which wastaken on directly.

The residue e from above was dissolved in H₂O (10 mL), AcOH (30 mL), THF(5 mL), and dichloroacetic acid (3 mL) and maintained at rt overnight.Water (5 mL) was added and the solution and maintained until hyrolysiswas complete, as monitored by HPLC-MS. Solid Na₂CO₃ was added cautiouslyuntil gas evolution ceased, the mixture was diluted with aq NaHCO₃, andextracted with 10% EtOAc/DCM. The combined organic phases were washedonce with brine, dried (Na₂SO₄), filtered, and concentrated. The residuewas purified by chromatography to afford 2.9 g of product f.

A mixture of ketone f (1.5 g) MeOH (50 ml) was treated with NaBH4 (290mg) at 0° C. for 20 min. The mixture was acidified to ˜pH1 with 10% aqcitric acid and the MeOH was removed under reduced pressure. The residuewas diluted with water and extraced with 20% EtOAc/DCM. The combinedorganic phases were washed once with brine, dried (Na₂SO₄), filtered,and concentrated. The residue was purified by chromatography to afford1.17 g of product g and 0.23 g of product h.

A mixture of ester g (1.17 g) LiOH.H2O (160 mg), THF (3 mL) and water(4.5 mL) was stirred vigorously at rt overnight. The mixture was dilutedwith brine and exhaustively extraced with EtOAc. The combined organicphases were washed once with brine, dried (Na₂SO₄), filtered, andconcentrated to afford acid i (525 mg).

Example 106 N-Boc-N-cyclopropylmethyl-L-alanine

L-alanine methyl ester hydrochloride a (5 g, 35.8 mmol) andcyclopropanecarboxaldehyde b (2.67 ml, 35.8 mmol) were suspended in 50ml THF w/1% AcOH. Addition of 5 ml of CH₃OH made the cloudy solutionturned to clear. NaCNBH₄ (2.25 g, 35.8 mmol) was added and the reactionmixture stirred overnight. The reaction was quenched by addition of 1Naq. NaOH, extracted by EtOAc twice, organic layers were dried overNa₂SO₄ and concentrated to dryness. The crude material was purified bychromatography using 30% EtOAc/hexane (stained by ninhydrin) to obtainthe compound c (1 g, 18%). The compound c (1 g, 6.37 mmol) anddi-t-bocdicarbonate (2.1 g, 9.55 mmol) were diluted in THF (20 ml) andH₂O (20 ml), NaHCO₃ (1.3 g, 15.9 mmol) was added. The reaction mixturestirred overnight for completion. THF was removed under reducedpressure, and the aqueous layer was extracted by EtOAc 3 times. Combinedorganic layers were washed by 1N NaOH, sat, NH₄Cl followed by brine, theconcentrated to dryness. The Boc-protected compound d (1.39 g, 5.40mmol) was stirred with LiOH.H₂O (1.14 g, 27 mmol) in THF (20 ml) and H₂O(20 ml) overnight at room temperature. THF was stripped off, and theaqueous layer was adjusted to pH=4 by adding 10% citric acid, thenextracted by EtOAc 3 times. Combined organic layers were washed by brineand concentrated. The crude was purified by reverse phase C-18 columneluted by 0%-50% acetonitrile/H₂O to give pure compound e as a whitesolid (794 mg).

Example 107

Phosphonate b (7.2 g, 21 mmol) was dissolved in THF (25 mL) at roomtemperature, and TMG (3.6 mL, 29 mmol, 1.3 equiv) was added dropwise.The mixture was stirred for 15 min at room temp. Commercially availableketone a (6.7 g, 43 mmol) was dissolved in THF (25 mL) and addeddropwise to the mixture of phosphonate and base. The reaction wasstirred for 24 h at room temperature and quenched by adding approx 200mL of 1 N HCl. Organic products were quickly extracted into 80% ethylacetate-hexanes (400 mL total). The combined organic phases were dried(Na₂SO₄), adsorbed onto Celite and purified twice by chromatography ISCOCombiFlash 120 g column, 0-55% ethyl acetate-hexanes over 20 min,followed by 55% ethyl acetate-hexanes for 5 min, to afford 3.83 g (10.6mmol, 50%) of the product amino ester c as a white solid.

Example 108

Ketal a (1.56 g, 4.73 mmol) was dissolved in 6 mL of THF. To thissolution was added deionized water (15 mL), glacial acetic acid (6 mL),and dichloroacetic acid (1 mL). The mixture was stirred overnight atroom temperature. Aqueous 1 N sodium hydroxide (approx. 100 mL) wasadded, and crude product was extracted into dichloromethane (approx. 200mL). The organic product was adsorbed onto Celite by evaporation of thesolvent, and purified by chromatography ISCO CombiFlash 80 g column witha solvent gradient of 0-40% ethyl acetate-hexanes over 20 min to afford452 mg (1.58 mmol, 33%) of ketone b.

Example 109

Ester a (184 mg, 0.55 mmol) was dissolved in 2 mL of THF. Deionizedwater was added (1 mL), followed by lithium hydroxide monohydrate (42mg, 1.0 mmol). The mixture was stirred at room temperature overnight,then acidified using aqueous 1 N HCl and extracted into dichloromethane.Drying (Na₂SO₄), filtration and evaporation of the solvent yielded 175mg (quantitative yield) of the carboxylic acid b.

Example 110

A small vial was charged with amine b (130 mg, 0.46 mmol), acid a (175mg, 0.55 mmol) and EDC.HCl (135 mg, 0.70 mmol). The mixture wasdissolved in dichloromethane (3 mL) and stirred overnight at roomtemperature. Celite was added to the reaction, and solvent was removedunder reduced pressure. Crude product was purified by chromatographyISCO CombiFlash 40 g column with a solvent gradient of 0-45% ethylacetate-hexanes over 10 min followed by 45% ethyl acetate-hexanes for 5min. The BOC-protected amine obtained from this coupling reaction wasdissolved in dichloromethane (2 mL), deionized water (0.5 mL) andtrifluoroacetic acid (1 mL) and allowed to stir for 3 h at roomtemperature. Organic solvents were removed under reduced pressure, theaqueous layer was made basic using a small amount of 1 N NaOH, andproduct was extracted into dichloromethane. Removal of organic solventyielded 110 mg (0.25 mmol, 45% amine #) of the free amine #.

Example 111

Standard EDC coupling procedure was performed using amine b (110 mg,0.25 mmol) L-BOC—N-methylalanine a (72 mg, 0.35 mmol) and EDC (67 mg,0.35 mmol). BOC-protected final product was purified by chromatographyISCO CombiFlash 12 g column with a solvent gradient of 5-55% ethylacetate-dichloromethane over 15 min followed by 55% ethylacetate-dichloromethane for 4 min. BOC-deprotection was performed using2:1 DCM:TFA with few drops of water. Final product c (54 mg, 66%) waspurified by reverse-phase HPLC C₁₈ column with a solvent gradient of5-50% acetonitrile-water over 20 min.

Example 112

Following the general procedure of Burk [Burk, M. J.; Gross, M. F.;Martinez, J. P. J. Am. Chem. Soc. 1995, 117, 9375-9376.], 5.0 g (13.8mmol) of alkene a, 100 mL of dry methanol, and[(S,S)-Me-BPE-Rh(COD)]⁺OTf⁻ (1.5 g, 2.4 mmol) were mixed in a Parrshaker flask purged with nitrogen. Parr shaker was evacuated andsubsequently charged to 70 psi of hydrogen gas for 32 hours. Methanolwas removed under reduced pressure, and crude product was filteredthrough a small plug of silica gel using ethyl acetate. Evaporation ofthe solvent gave 4.0 g (11 mmol, 80%) of product b with >98% yield.

Example 113

Z-protected amino ester a (4.0 g, 11 mmol) was dissolved in methanol (30mL). To this solution was added BOC-anhydride (2.9 g, 13.5 mmol),followed by 20% Pd(OH)₂.C (1.0 g). All air was removed from the reactionflask by house vacuum, and the mixture was stirred vigorously for 5 min.The flask was then filled with hydrogen gas and allowed to stirvigorously at room temperature for 6 h. After evacuating the hydrogenatmosphere, the mixture was filtered through Celite using methanol, andcrude product was obtained by evaporation of the solvent.

The product BOC-protected amine b was dissolved in 5 mL of THF. Thefollowing solvents were then added sequentially: deionized water (15mL), glacial acetic acid (30 mL), and dichloroacetic acid (3 mL). Themixture was stirred overnight at room temperature, and the reaction wasquenched by slowly adding solid sodium carbonate with vigorous stirringuntil the release of gas was no longer visible. Crude product wasextracted into 10% ethyl acetate-dichloromethane. The product wasadsorbed onto Celite by evaporation of the solvents, and purified bychromatography ISCO CombiFlash 120 g column with a solvent gradient of0-36% ethyl acetate-hexanes over 20 min followed by flushing with 36%ethyl acetate-hexanes for 5 min to afford 2.86 g (10.0 mmol, 91%) ofketone b.

Example 114

Standard EDC coupling was performed using amine b (46 mg, 0.15 mmol),carboxylic acid a (42 mg, 0.15 mmol syn-diastereomer) and EDC (33 mg,0.17 mmol). BOC-protected final product was purified by chromatographyISCO CombiFlash stacker 2×4 g column with a solvent gradient of 0-28%ethyl acetate-dichloromethane over 15 min, followed by 28% ethylacetate-dichloromethane for 3 min. Standard BOC-deprotection wasperformed using 2:1 DCM:TFA with few drops of water. The TFA salt wastreated with base (aqueous 1 N NaOH) and extracted into dichloromethane.

Example 115

Coupling of the product primary amine a (20 mg, 0.044 mmol) toL-BOC—N-methylalanine b (12 mg, 0.059 mmol) was performed by adding EDC(10 mg 0.052 mmol) and dissolving in dichloromethane (1 mL).BOC-protected final product was purified by chromatography ISCOCombiFlash stacker 2×12 g column with a solvent gradient of 0-70% ethylacetate-dichloromethane over 20 min followed by 70% ethylacetate-dichloromethane for 5 min. BOC-deprotection was performed using2:1 DCM:TFA with few drops of water. Final product c was purified byreverse-phase HPLC C₁₈ column with a gradient of 5-50%acetonitrile-water over 20 min. Yield of product anti-diastereomer c was22 mg.

Example 116

Standard EDC coupling was performed using amine b (110 mg, 0.38 mmol),carboxylic acid a, (105 mg, 0.38 mmol) and EDC (86 mg, 0.45 mmol).BOC-protected final product was purified by chromatography ISCOCombiFlash stacker 2×4 g column with a solvent gradient of 0-28% ethylacetate-dichloromethane over 15 min, followed by 28% ethylacetate-dichloromethane for 3 min. Standard BOC-deprotection wasperformed using 2:1 DCM:TFA with few drops of water. The TFA salt wastreated with base (aqueous 1 N NaOH) and extracted into dichloromethane.

Example 117

Coupling of the product primary amine b (170 mg, 0.35 mmol) toL-BOC—N-methylalanine a (81 mg, 0.40 mmol) was performed by adding EDC(77 mg 0.40 mmol) and dissolving in dichloromethane (2 mL).BOC-protected final product was purified by chromatography ISCOCombiFlash stacker 2×12 g column with a solvent gradient of 0-70% ethylacetate-dichloromethane over 20 min followed by 70% ethylacetate-dichloromethane for 5 min. Standard BOC-deprotection wasperformed using 2:1 DCM:TFA with few drops of water. Final product c waspurified by reverse-phase HPLC C₁₈ column with a solvent gradient of5-50% acetonitrile-water over 20 min. Yield of anti-diastereomer productc was 106 mg.

Example 118

Ketone a (1.45 g, 5.3 mmol), was dissolved in dry diethyl ether (20 mL)and cooled to −78° C. Methyllithium (1.6 M in Et₂O, 9.5 mL, 15 mmol) wasadded dropwise to the reaction mixture and stirred vigorously at thereduced temperature for 1 h. The reaction was quenched by pouring thecold mixture into saturated aqueous ammonium chloride and extracting theorganics into dichloromethane. The organic layer was dried (Na₂SO₄),filtered, adsorbed onto Celite and purified by chromatography ISCOCombiFlash 120 g column, 0-50% ethyl acetate-hexanes over 25 min,followed by flushing 50% ethyl acetate-hexanes for 3 min, and 90% ethylacetate-hexanes for 3 min. This purification afforded 344 mg (1.1 mmol,42%) of the syn-diastereomer c and 299 mg (0.99 mmol, 37%) of theanti-diastereomer b.

Example 119

Hydrolysis of the methyl ester a (300 mg, 0.99 mmol) was carried out bydissolving in THF (0.8 mL), adding deionized water (1.2 mL) and LiOH.H₂O(47 mg, 1.1 mmol). The mixture was stirred at room temperature for 2 h,then reacidified using aqueous 1 N HCl and extracted into 90% ethylacetate-dichloromethane. Brine was added to the aqueous acid layer toaid in the extraction. Drying (Na₂SO₄), filtration, and evaporation ofthe solvent yielded the carboxylic acid b (79 mg, 0.28 mmol).

Example 120

Hydrolysis of the methyl ester a (340 mg, 1.1 mmol) was carried out bydissolving in THF (0.9 mL), adding deionized water (1.4 mL) and LiOH.H₂O(50 mg, 1.2 mmol). The mixture was stirred at room temperature for 2 h,then reacidified using aqueous 1 N HCl and extracted into 90% ethylacetate-dichloromethane. Brine was added to the aqueous acid layer toaid in the extraction. Drying (Na₂SO₄), filtration, and evaporation ofthe solvent yielded the carboxylic acid b (254 mg, 0.88 mmol), cleanenough to use in the next step without purification.

Example 121

Standard EDC coupling was performed using amine b (62 mg, 0.21 mmol),the carboxylic acid a, (32 mg, 0.11 mmol) and EDC (21 mg, 0.11 mmol).BOC-protected final product was purified by chromatography ISCOCombiFlash 12 g column with a solvent gradient of 0-40% ethylacetate-dichloromethane over 22 min, followed by 67% ethylacetate-dichloromethane for 3 min. Standard BOC-deprotection wasperformed using 2:1 DCM: TFA with few drops of water. The TFA salt wastreated with base (aqueous 1 N NaOH) and extracted into ethyl acetatewith 10% dichloromethane.

Example 122

Coupling of the primary amine b (47 mg, 0.1 mmol) toL-BOC—N-methylalanine a (65 mg, 0.30 mmol) was performed by adding EDC(61 mg, 0.32 mmol) and dissolving in dichloromethane (2 mL).BOC-protected final product was purified by chromatography ISCOCombiFlash 12 g column with a solvent gradient of 5-65% ethylacetate-dichloromethane over 25 min. Standard BOC-deprotection wasperformed using 2:1 DCM:TFA+few drops of water. Final product c waspurified by reverse-phase HPLC C₁₈ column with a solvent gradient of5-50% acetonitrile-water over 20 min. Yield of anti-diastereomer productc was 22 mg (31% from proline amine starting material).

Example 123

Standard EDC coupling was performed using amine b (82 mg, 0.27 mmol),the carboxylic acid a, (95 mg, 0.33 mmol) and EDC (65 mg, 0.34 mmol).BOC-protected final product was purified by chromatography ISCOCombiFlash 12 g column with a solvent gradient of 0-40% ethylacetate-dichloromethane over 22 min, followed by 67% ethylacetate-dichloromethane for 3 min. Standard BOC-deprotection wasperformed using 2:1 DCM:TFA with few drops of water. The TFA salt wastreated with base (aqueous 1 N NaOH) and extracted into ethyl acetatewith 10% dichloromethane.

Example 124

Coupling of the product primary amine b (70 mg, 0.15 mmol) toL-BOC—N-methylalanine a (37 mg, 0.18 mmol) was accomplished by addingEDC (36 mg 0.19 mmol) and dissolving in dichloromethane (2 mL).BOC-protected final product was purified by chromatography ISCOCombiFlash 12 g column with a solvent gradient of 1-51% ethylacetate-dichloromethane over 20 min followed by 51% ethylacetate-dichloromethane for 3 min. Standard BOC-deprotection wasperformed using 2:1 DCM:TFA+few drops of water. Final product b waspurified by reverse-phase HPLC C₁₈ column with a solvent gradient of5-50% acetonitrile-water over 20 min. Yield of product b was 49 mg.

Example 125

Sulfide a (810 mg, 2.5 mmol), synthesized according to the generalprocedure of Shieh [Shieh, W-C.; Xue, S.; Reel, N.; Wu, R.; Fitt, J.;Repic, O. Tetrahedron: Asymmetry, 2001, 12, 2421-2425], was dissolved inmethanol (25 mL). Oxone (4.5 g) was dissolved in deionized water (25mL). The methanol solution of substrate was cooled to −10° C., and theaqueous solution of Oxone was added to the reaction slowly. The reactionwas kept on ice and gradually allowed to warm to room temperature whilestirring overnight. Deionized water was used to dilute the reaction toapprox 150 mL, which was poured into 90% ethyl acetate-hexanes forextraction. The organic phase was dried (Na₂SO₄), adsorbed onto Celiteand purified by chromatography ISCO CombiFlash 40 g column, 5-90% ethylacetate-hexanes over 30 min to afford 804 mg (2.27 mmol, 91%) of theproduct sulfone b.

Example 126

Following the general procedure of Burk [Burk, M. J.; Gross, M. F.;Martinez, J. P. J. Am. Chem. Soc. 1995, 117, 9375-9376.], alkene a (774mg 2.19 mmol), dry methanol (40 mL), and [(S,S)-Me-BPE-Rh(COD)]⁺OTf⁻(500 mg, 0.8 mmol) were mixed in a Parr shaker flask purged withnitrogen. Parr shaker was evacuated and subsequently charged to 60 psiof hydrogen gas and shaken vigorously overnight. Methanol was removedunder reduced pressure, and crude product was filtered through a smallplug of silica gel using ethyl acetate. Evaporation of the solventyielded 730 mg (2.0 mmol, 94%) of product b with >98% yield.

Example 127

Z-protected amino ester a (804 mg, 2.27 mmol) was dissolved in methanol(16 mL). To this solution was added BOC-anhydride (1.5 g, 6.8 mmol),followed by 20% Pd(OH)₂.C (250 mg). All air was removed from thereaction flask by house vacuum, and the mixture was stirred vigorouslyfor 5 min. The flask was then filled with hydrogen gas and allowed tostir vigorously at room temperature for 6 h. After evacuating thehydrogen atmosphere, the mixture was filtered through Celite usingmethanol, and crude product b was obtained by evaporation of the solvent(508 mg, 1.56 mmol, 70% yield).

Example 128

Ester a (508 mg, 1.56 mmol) was dissolved in 8 mL of THF. Deionizedwater (4 mL) was added, followed by LiOH.H₂O (120 mg, 2.8 mmol). Themixture was stirred at room temperature overnight, acidified usingaqueous 1 N HCl and extracted into ethyl acetate. Drying (Na₂SO₄),filtration and evaporation of the solvent yielded 372 mg (1.21 mmol, 78%yield) of the carboxylic acid b, clean enough to use in the next stepwithout purification.

Example 129

Standard EDC coupling was performed using amine b (100 mg, 0.2 mmol),the carboxylic acid a, (58 mg, 0.29 mmol) and EDC (56 mg, 0.29 mmol).BOC-protected final product was purified by chromatography ISCOCombiFlash 12 g column with a solvent gradient of 0-65% ethylacetate-dichloromethane over 15 min. Standard BOC-deprotection wasperformed using 2:1 DCM:TFA with few drops of water. Final product c waspurified by reverse-phase HPLC C₁₈ column with a solvent gradient of5-50% acetonitrile-water over 18 min. Yield of product c was 132 mg.

Example 130

Standard EDC coupling was performed using amine b (130 mg, 0.3 mmol),the carboxylic acid a, (60 mg, 0.28 mmol) and EDC (60 mg, 0.3 mmol).BOC-protected final product was purified by chromatography ISCOCombiFlash 12 g column with a solvent gradient of 0-65% ethylacetate-dichloromethane over 15 min. Standard BOC-deprotection wasperformed using 2:1 DCM:TFA with few drops of water. Final product c waspurified by reverse-phase HPLC C₁₈ column with a solvent gradient of5-50% acetonitrile-water over 18 min. Yield of product c was 78 mg.

Example 131

Following the general procedure of Grigg [Blaney, P.; Grigg, R.;Rankovic, Z.; Thornton-Pett, M.; Xu, J. Tetrahedron, 2002, 58,1719-1737] a roundbottom flask was charged with sodium hydride (480 mg60% dispersion in oil, 12.0 mmol, 4.0 equiv) and purged with nitrogenfor 15 min. THF (6.0 mL) was added to the flask, and the suspension wascooled to 0° C. using an ice water bath. A separate flask was chargedwith BOC-glycine a (525 mg, 3.0 mmol), dry THF (6.0 mL) and ethyl iodide(1.0 mL, 12 mmol, 4 equiv). This mixture was added dropwise to the NaHsuspension in THF, with vigorous stirring at 0° C. After 1 h ofstirring, the reaction was warmed to room temperature and allowed tostir overnight. The reaction was again cooled to 0° C., and methanol (4mL) was added very slowly to quench the excess hydride. Deionized waterwas added to dilute the mixture, and methanol was removed under reducedpressure. Impurities were extracted into 90% ethyl acetate-hexanes, theaqueous layer was then acidified by adding solid citric acid until thepH reached 2-3. The product was extracted into 90% ethylacetate-hexanes. This organic layer was dried (Na₂SO₄) and filtered.Removal of the solvents under reduced pressure afforded a quantitativeyield of the product b.

Example 132

Standard EDC coupling was performed using amine b (70 mg, 0.16 mmol),the carboxylic acid a, (49 mg, 0.24 mmol) and EDC (46 mg, 0.24 mmol).BOC-protected final product was purified by chromatography ISCOCombiFlash 12 g column with a solvent gradient of 0-55% ethylacetate-dichloromethane over 15 min. Standard BOC-deprotection wasperformed using 2:1 DCM:TFA with few drops of water. Final product c waspurified by reverse-phase HPLC C₁₈ column with a solvent gradient of5-50% acetonitrile-water over 18 min. Yield of product c was 82 mg.

Example 133

The free primary amine b (35 mg, 0.056 mmol), anhydrous potassiumcarbonate (70 mg, 0.5 mmol) and formamidine hydrochloride a (30 mg, 0.37mmol) were mixed together in a vial and dissolved in methanol (1.2 mL).The mixture was stirred at room temperature for 1.5 hr. Glacial aceticacid was added until gas release was no longer visible, and the mixturewas filtered. Reverse-phase HPLC, using a Cl₈ column and a solventgradient of 5-50% acetonitrile-water over 25 min with 0.1% TFA,separated the desired product c, affording 8.2 mg (0.015 mmol, 27%yield) of the TFA salt after lyophilization.

Example 134

A mixture of unprotected amino acid a (775 mg, 7.24 mmol) and sodiumcarbonate (1.69 g, 16.0 mmol) was dissolved in a 1:1 solution ofdeionized water and THF (15 mL each). To this mixture was addedBOC-anhydride b (1.73 g, 7.96 mmol). The mixture was stirred at roomtemperature overnight, and THF was removed under reduced pressure. Themixture was then acidified to pH 2-3 with saturated aqueous citric acid,and product was extracted into 10% ethyl acetate-dichloromethane. Theorganic layer was dried (Na₂SO₄), filtered and concentrated underreduced pressure to afford clean BOC-protected amino acid c (1.40 g, 6.7mmol, 93%) to be used without further purification.

Example 135

Standard EDC coupling was performed using amine b (64 mg, 0.14 mmol),the carboxylic acid a, (41 mg, 0.2 mmol) and EDC (38 mg, 0.2 mmol).BOC-protected final product was purified by chromatography ISCOCombiFlash 12 g column with a solvent gradient of 0-55% ethylacetate-dichloromethane over 10 min, followed by a steady flow of 55%ethyl acetate-dichloromethane for 3 min. Standard BOC-deprotection wasperformed using 2:1 DCM:TFA+few drops of water. Final product c waspurified by reverse-phase HPLC C₁₈ column with a solvent gradient of5-50% acetonitrile-water over 18 min. Yield of product c was 70.2 mg.

Example 136

Standard EDC coupling was performed using amine hydrochloride b (250 mg,0.67 mmol), the carboxylic acid a, (187 mg, 0.81 mmol), DIPEA (0.35 mL,2.0 mmol) and EDC (157 mg, 0.81 mmol). Reaction was stirred at roomtemperature for 48 h. BOC-protected final product was purified bychromatography ISCO CombiFlash 12 g column with a solvent gradient of0-25% ethyl acetate-hexanes over 10 min, followed by a steady flow of26% ethyl acetate-hexanes for 3 min. Standard BOC-deprotection wasperformed using HCl in dioxane (4.0 M, 3.0 mL).

To the primary amine hydrochloride c (170 mg, 0.38 mmol) andL-BOC—N-methylalanine (91 mg, 0.45 mmol), was added dichloromethane (2mL), DIPEA (0.20 mL, 1.1 mmol) and EDC (86 mg, 0.45 mmol), stirring atroom temperature for 24 h. BOC-protected final product was purified bychromatography ISCO CombiFlash 12 g column with a solvent gradient of0.5-52% ethyl acetate-hexanes over 13 min followed by 52% ethylacetate-hexanes for 3 min. Standard BOC-deprotection was performed using2:1 DCM:TFA with few drops of water. Final product was purified byreverse-phase HPLC C₁₈ column with a solvent gradient of 5-60%acetonitrile-water over 20 min. Yield of final product was 90 mg.

Example 137

Standard EDC coupling was performed using amine hydrochloride # (250 mg,0.67 mmol), the carboxylic acid a, (187 mg, 0.81 mmol), DIPEA (0.350 mL,2.0 mmol) and EDC (157 mg, 0.81 mmol). Reaction was stirred at roomtemperature for 3 h. BOC-protected final product was purified bychromatography ISCO CombiFlash 12 g column with a solvent gradient of0-25% ethyl acetate-hexanes over 10 min, followed by a steady flow of26% ethyl acetate-hexanes for 3 min. Standard BOC-deprotection wasperformed using HCl in dioxane (4.0 M, 3.0 mL).

To the primary amine hydrochloride (160 mg, 0.35 mmol) andL-BOC—N-methylalanine (91 mg, 0.45 mmol), was added dichloromethane (2mL), DIPEA (0.200 mL, 1.1 mmol) and EDC (86 mg, 0.45 mmol), stirring atroom temperature for 24 h. BOC-protected final product was purified bychromatography ISCO CombiFlash 12 g column with a solvent gradient of0.5-52% ethyl acetate-hexanes over 13 min followed by 52% ethylacetate-hexanes for 3 min. Standard BOC-deprotection was performed using2:1 DCM:TFA with few drops of water. Final product was purified byreverse-phase HPLC C₁₈ column with a solvent gradient of 5-60%acetonitrile-water over 20 min. Yield of product c was 79 mg.

Example 138

EDC coupling was performed using amine hydrochloride b (230 mg, 0.61mmol), the carboxylic acid a, (165 mg, 0.75 mmol), DIPEA (0.350 mL, 2.0mmol) and EDC (157 mg, 0.81 mmol). Reaction was stirred at roomtemperature for 3 h, LC/MS indicated only half complete. More carboxylicacid (160 mg) and EDC (150 mg) was added to the reaction, and themixture was stirred overnight at room temperature. BOC-protected finalproduct was purified by chromatography ISCO CombiFlash 40 g column witha solvent gradient of 0-55% ethyl acetate-hexanes over 17 min, followedby a steady flow of 56% ethyl acetate-hexanes for 5 min. StandardBOC-deprotection was performed using 2:1 DCM:TFA+few drops of water.Coupling of the product primary amine c (199 mg, 0.5 mmol) toL-BOC—N-methylalanine (140 mg, 0.7 mmol) was performed with EDC (135 mg,0.7 mmol) and dichloromethane (3 mL). BOC-protected final product waspurified by chromatography ISCO CombiFlash 12 g column with a solventgradient of 0-40% ethyl acetate-dichloromethane over 15 min followed by40% ethyl acetate-dichloromethane for 3 min. Standard BOC-deprotectionwas performed using 2:1 DCM:TFA with few drops of water. Final productwas purified by reverse-phase HPLC C₁₈ column with a solvent gradient of5-50% acetonitrile-water over 20 min. Yield of final product was 178 mg.

Example 139

Methyl ketone a (480 mg, 3.0 mmol), synthesized according to the generalprocedure of Miki [Miki, Y.; Nakamura, N.; Hachiken, H.; Takemura, S. J.Heterocyclic Chem., 1989, 26, 1739-1745], was suspended in 33% HBr inacetic acid (6 mL). Elemental bromine was added in six portions (6×0.025mL, 0.15 mL total, 3.0 mmol) with vigorous stirring at room temperature.The reaction appeared to have a light color after 10 min of stirring,when diethyl ether was added (10 mL). Stirring at room temperature wascontinued for 30 min. The mixture was filtered through a frit, and thesolids left behind were rinsed with 20 mL of ether, transferred to avial, and dried under high vacuum. The solid afforded (840 mg) was amixture of desired product b and the HBr salt of the starting material,used without further purification in the thiazole-forming step.

Example 140

Thioamide a (2.26 mg, 9.8 mmol) was added to the mixture of bromomethylketone b and methyl ketone (1.44 g) in a roundbottom flask. Ethanol (30mL) was added, dissolving the thioamide and suspending the salts.Pyridine was then added dropwise (0.4 mL, 5.0 mmol) and the mixture wasstirred at room temperature for 5 min. The reaction flask was thenheated to 70° C. in an oil bath, with vigorous stirring. After 10 min,the suspension of salts was no longer visible and the reaction washomogeneous. The reaction was allowed to cool to room temperature for 45min, and Celite was added along with toluene (20 mL). Solvents wereremoved under reduced pressure. The crude product adsorbed onto Celitewas purified by chromatography ISCO CombiFlash 120 g column, 0-30% ethylacetate-dichloromethane over 20 min, followed by a gradient of 30-70%ethyl acetate-dichloromethane over 5 min, to afford 518 mg (1.4 mmol,47%) of the product thiazole. Removal of BOC from the proline amine wasaccomplished by dissolving the substrate in 2:1 DCM:TFA with few dropsof water, following the standard procedure. Free base was obtained bytreating the TFA salt with 1 N aqueous sodium hydroxide and extractingthe amine into dichloromethane. Drying of the organic layer (Na₂SO₄),filtering and removing the solvent under reduced pressure afforded 356mg (1.3 mmol, 93%) of free amine c.

Example 141

HOAt, DIC procedure was used to couple the above dipeptide to amine.Secondary amine b (65 mg, 0.25 mmol), carboxylic acid a, (97 mg, 0.28mmol) HOAt (53 mg, 0.4 mmol) and DIC (50 mg, 0.4 mmol). BOC-protectedfinal product was purified by chromatography ISCO CombiFlash 12 g columnwith a solvent gradient of 0-65% ethyl acetate-dichloromethane over 15min. Standard BOC-deprotection was performed using 2:1 DCM:TFA+few dropsof water. Final product c was purified by reverse-phase HPLC C₁₈ columnwith a solvent gradient of 5-50% acetonitrile-water over 18 min. Yieldof product c was 98 mg.

Example 142

HOAt, DIC procedure was used to couple the above dipeptide to amine.Secondary amine b (50 mg, 0.2 mmol), carboxylic acid a, (72 mg, 0.21mmol) HOAt (40 mg, 0.3 mmol) and DIC (38 mg, 0.3 mmol). BOC-protectedfinal product was purified by chromatography ISCO CombiFlash 12 g columnwith a solvent gradient of 10-85% ethyl acetate-dichloromethane over 20min. Standard BOC-deprotection was performed using 2:1 DCM:TFA+few dropsof water. Final product c was purified by reverse-phase HPLC C₁₈ columnwith a solvent gradient of 3-40% acetonitrile-water over 20 min. Yieldof final product c was 25 mg.

Example 143

A mixture of unprotected amino acid a (1.1 g, 10 mmol) and sodiumcarbonate (850 mg, 10 mmol) was dissolved in a 1:1 solution of deionizedwater and THF (13 mL each). To this mixture was added FMOC-OSu b (6×550mg, total 3.3 g, 9.8 mmol) over a period of 1 h. After each addition ofFMOC-OSu was added 2-3 mL of 1 M aqueous sodium bicarbonate to keep thereaction mixture at basic pH. The mixture was stirred at roomtemperature overnight, and THF was removed under reduced pressure. Themixture was then diluted with deionized water, poured into ethyl acetatein a separatory funnel, and made acidic by the addition of 6 N HCl.After extracting into ethyl acetate, the organic layer was washed withdeionized water, followed by brine. The organic layer was dried(Na₂SO₄), filtered and concentrated under reduced pressure to affordclean FMOC-protected amino acid c (1.05 g, 3.19 mmol, 32%) to be usedwithout further purification.

Example 144

Following the general procedure of Freidinger [Freidinger, R. M.;Hinkle, J. S.; Perlow, D. S.; Arison, B. H. J. Org. Chem., 1983, 48,77-81], the FMOC-protected primary amine a (1.04 g, 3.17 mmol) wasdissolved in toluene (60 mL). Paraformaldehyde (630 mg) was added,followed by a catalytic amount of p-toluenesulfonic acid (70 mg, 0.37mmol). The mixture was vigorously stirred at reflux temperature for 45min, collecting any generated water in a Dean Stark trap. The reactionmixture was then allowed to cool to room temperature, and washed withsaturated aqueous sodium bicarbonate (2×30 mL). The organic layer wasdried (Na₂SO₄), filtered, and concentrated under reduced pressure toafford 910 mg (2.7 mmol) of oxazolidinone b. The oxazolidinone (337 mg,0.99 mmol) was dissolved in dichloromethane (20 mL). To this solutionwas added anhydrous aluminum trichloride (260 mg, 2.0 mmol), followed bytriethylsilane (0.32 mL, 2.0 mmol). The reaction mixture was stirred for5 h at room temperature and then quenched with 20 mL of 1 N aqueous HCl.The product carboxylic acid was extracted into 25% ethylacetate-dichloromethane and washed with 1 N aqueous HCl (20 mL) followedby brine. The organic layer was dried (Na₂SO₄) and filtered. Celite wasadded, and the solvent was removed under reduced pressure. The crudeproduct adsorbed onto Celite was purified by chromatography ISCOCombiFlash 40 g column, 1-55% ethyl acetate-dichloromethane over 25 min,to afford 272 mg (0.79 mmol, 25% yield from FMOC-primary amine) of theFMOC protected N-methyl amino acid c.

Example 145

Standard EDC coupling was performed using amine # (140 mg, 0.4 mmol),the crude carboxylic acid a, (176 mg, 0.4 mmol) and EDC (80 mg, 0.4mmol). BOC-protected final product was purified by chromatography ISCOCombiFlash 40 g column with a solvent gradient of 1-40% ethylacetate-dichloromethane over 20 min. The desired BOC-protected productwas split into two portions for removing the FMOC group. The firstportion (50 mg, 0.065 mmol) was dissolved in dichloromethane (1.0 mL),treated with piperidine (0.10 mL, 1.0 mmol), and allowed to stir at roomtemperature for 2 h. The second portion (100 mg, 0.13 mmol) wasdissolved in 20% piperidine in DMF (1.0 mL) and allowed to stir at roomtemperature overnight. Both reactions were quenched by adding few dropsof TFA. Final product was purified by reverse-phase HPLC C₁₈ column witha solvent gradient of 3-40% acetonitrile-water over 20 min. Combinedyield of final product c was 55 mg.

Example 146 IAP Inhibition Assays

In the following experiments was used a chimeric BIR domain referred toas MLXBIR3SG in which 11 of 110 residues correspond to those found inXIAP-BIR3, while the remainder correspond to ML-IAP-BIR. The chimericprotein MLXBIR3SG was shown to bind and inhibit caspase-9 significantlybetter than either of the native BIR domains, but bound Smac-basedpeptides and mature Smac with affinities similar to those of nativeML-IAP-BIR. The improved caspase-9 inhibition of the chimeric BIR domainMLXBIR3SG has been correlated with increased inhibition ofdoxorubicin-induced apoptosis when transfected into MCF7 cells.

MLXBIR3SG sequence: (SEQ ID NO.: 1)MGSSHHHHHHSSGLVPRGSHMLETEEEEEEGAGATLSRGPAFPGMGSEELRLASFYDWPLTAEVPPELLAAAGFFHTGHQDKVRCFFCYGGLQSWKRGDDPWTEHAKWFPGCQFLLRSKGQEYINNIHLTHSL

TR-FRET Peptide Binding Assay

Time-Resolved Fluorescence Resonance Energy Transfer competitionexperiments were performed on the Wallac Victor2 Multilabeled CounterReader (Perkin Elmer Life and Analytical Sciences, Inc.) according tothe procedures of Kolb et al (Journal of Biomolecular Screening, 1996,1(4):203). A reagent cocktail containing 300 nM his-tagged MLXBIR3SG;200 nM biotinylated SMAC peptide (AVPI); 5 μg/mL anti-hisallophycocyanin (XL665) (CISBio International); and 200 ng/mLstreptavidin-europium (Perkin Elmer) was prepared in reagent buffer (50mM Tris [pH 7.2], 120 mM NaCl, 0.1% bovine globulins, 5 mM DTT and 0.05%octylglucoside). (Alternatively, this cocktail can be made usingeuropium-labeled anti-His (Perkin Elmer) andstreptavidin-allophycocyanin (Perkin Elmer) at concentrations of 6.5 nMand 25 nM, respectively). The reagent cocktail was incubated at roomtemperature for 30 minutes. After incubation, the cocktail was added to1:3 serial dilutions of an antagonist compound (starting concentrationof 50 μM) in 384-well black FIA plates (Greiner Bio-One, Inc.). After a90 minute incubation at room temperature, the fluorescence was read withfilters for the excitation of europium (340 nm) and for the emissionwavelengths of europium (615 nm) and a allophycocyanin (665 nm).Antagonist data were calculated as a ratio of the emission signal ofallophycocyanin at 665 nm to that of the emission of europium at 615 nm(these ratios were multiplied by a factor of 10,000 for ease of datamanipulation). The resulting values were plotted as a function ofantagonist concentration and fit to a 4-parameter equation usingKaleidograph software (Synergy Software, Reading, Pa.). Indications ofantagonist potency were determined from the IC50 values. Compounds ofthe invention that were tested in this assay exhibited IC50 values ofless than 200 μM indicating IAP inhibitory activity.

Fluorescence Polarization Peptide Binding Assay

Polarization experiments were performed on an Analyst HT 96-384(Molecular Devices Corp.) according to the procedure of Keating, S. M.,Marsters, J, Beresini, M., Ladner, C., Zioncheck, K., Clark, K.,Arellano, F., and Bodary., S. (2000) in Proceedings of SPIE: In VitroDiagnostic Instrumentation (Cohn, G. E., Ed.) pp 128-137, Bellingham,Wash. Samples for fluorescence polarization affinity measurements wereprepared by addition of 1:2 serial dilutions starting at a finalconcentration of 5 μM of MLXBIR3SG in polarization buffer (50 mM Tris[pH 7.2], 120 mM NaCl, 1% bovine globulins 5 mM DTT and 0.05%octylglucoside) to 5-carboxyflourescein-conjugated AVPdi-Phe-NH₂(AVP-diPhe-FAM) at 5 nM final concentration.

The reactions were read after an incubation time of 10 minutes at roomtemperature with standard cut-off filters for the fluoresceinfluorophore (λ_(ex)=485 nm; λ_(em)=530 nm) in 96-well black HE96 plates(Molecular Devices Corp.). Fluorescence values were plotted as afunction of the protein concentration, and the IC50s were obtained byfitting the data to a 4-parameter equation using Kaleidograph software(Synergy software, Reading, Pa.). Competition experiments were performedby addition of the MLXBIR3SG at 30 nM to wells containing 5 nM of theAVP-diPhe-FAM probe as well as 1:3 serial dilutions of antagonistcompounds starting at a concentration of 300 μM in the polarizationbuffer. Samples were read after a 10-minute incubation. Fluorescencepolarization values were plotted as a function of the antagonistconcentration, and the IC₅₀ values were obtained by fitting the data toa 4-parameter equation using Kaleidograph software (Synergy software,Reading, Pa.). Inhibition constants (K_(i)) for the antagonists weredetermined from the IC₅₀ values. Compounds of the invention that weretested in this assay exhibited a Ki or less than 100 μM.

We claim:
 1. A compound of formula I:

wherein A is thiazole; ring A and Q together are selected from the groupconsisting of:

Q is (a) alkyl, wherein one or more CH₂ groups of an alkyl is optionallyreplaced with —O—, —S—, —S(O)—, —S(O)₂—, —N(R₈)—, C(O)—NR₈—, —NR₈—C(O)—,—SO₂—NR₈—, —NR₈—SO₂—, —NR₈—C(O)—NR₈—, —NR₈—C(NH)NR₈— or —NR₈—C(NH)—; andthe alkyl is optionally substituted with one or more halogen, cyano,nitro, optionally substituted carbocycle or optionally substitutedheterocycle; or (b) a carbocycle or heterocycle optionally substitutedwith one or more halogen, nitro, cyano, oxo, alkyl, carbocycle orheterocycle; wherein one or more CH₂ groups of the alkyl substituent onQ is optionally replaced with —O—, —S—, —S(O)—, —S(O)₂—, —N(R₈)—,—C(O)—, —C(O)—NR₈—, —NR₈—C(O)—, —SO₂—NR₈—, —NR₈—SO₂—, —NR₈—C(O)—NR₈—,—NR₈—C(NH)NR₈—, —NR₈—C(NH)—, —C(O)—O— or —O—C(O)—, and the alkyl isoptionally substituted with one or more halogen, cyano, nitro, anoptionally substituted carbocycle or an optionally substitutedheterocycle; and wherein the carbocycle or heterocycle substituents on Qare optionally substituted with one or more hydroxyl, alkoxy, acyl,halogen, mercapto, oxo, carboxyl, halo-substituted alkyl, amino, cyano,nitro, amidino, guanidino, carbocycle or heterocycle; X₁ and X₂ are eachindependently O or S; Y is CH₂; R₁ is H, or R₁ and R₂ together form a5-8 member ring; R₂ is alkyl, a carbocycle, carbocyclylalkyl, aheterocycle or heterocyclylalkyl, each optionally substituted withhalogen, hydroxyl, oxo, thione, mercapto, carboxyl, alkyl, haloalkyl,alkoxy, alkylthio, sulfonyl, amino or nitro; R₃ is H or alkyl optionallysubstituted with halogen or hydroxyl; or R₃ and R₄ together form a 3-6member heterocycle; R₃′ is H, or R₃ and R₃′ together form a 3-6 membercarbocycle; R₄ is H, hydroxyl, amino, alkyl, carbocycle,carbocycloalkyl, carbocycloalkyloxy, carbocycloalkyloxycarbonyl,heterocycle, heterocycloalkyl, heterocycloalkyloxy orheterocycloalkyloxycarbonyl; wherein each alkyl, carbocycloalkyl,carbocycloalkyloxy, carbocycloalkyloxycarbonyl, heterocycle,heterocycloalkyl, heterocycloalkyloxy and heterocycloalkyloxycarbonyl isoptionally substituted with halogen, hydroxyl, mercapto, carboxyl,alkyl, alkoxy, amino, imino, or nitro; or R₄ and R₄′ together form aheterocycle; R₄′ is H; R₅ is H or alkyl; R₆ and R₆′ are eachindependently H, alkyl, aryl or aralkyl; R₇ in each instance isindependently H, cyano, hydroxyl, mercapto, halogen, nitro, carboxyl,amidino, guanidino, alkyl, a carbocycle, a heterocycle or —U—V; whereinU is —O—, —S—, —S(O)—, —S(O)₂—, —N(R₈)—, —C(O)—, —C(O)—NR₈—, —NR₈—C(O)—,—SO₂—NR₈—, —NR₈—SO₂—, —NR₈—C(O)—NR₈—, —NR₈—C(NH)NR₈—, —NR₈—C(NH)—,—C(O)—O— or —O—C(O)—; and V is alkyl, a carbocycle or a heterocycle; andwherein one or more CH₂ groups of an alkyl is optionally replaced with—O—, —S—, —S(O)—, —S(O)₂—, —N(R₈)—, —C(O)—, —C(O)—NR₈—, —NR₈—C(O)—,—SO₂—NR₈—, —NR₈—SO₂—, —NR₈—C(O)—NR₈—, —NR₈—C(NH)NR₈—, —NR₈—C(NH)—,—C(O)—O— or —O—C(O)—; and an alkyl, carbocycle and heterocycle isoptionally substituted with hydroxyl, alkoxy, acyl, halogen, mercapto,oxo, carboxyl, halo-substituted alkyl, amino, cyano, nitro, amidino,guanidino, an optionally substituted carbocycle or an optionallysubstituted heterocycle; R₈ is H, alkyl, a carbocycle or a heterocycle;wherein one or more CH₂ groups of said alkyl is optionally replaced with—O, —S—, —S(O)—, —S(O)₂— or —C(O)—; and said alkyl, carbocycle andheterocycle is optionally substituted with hydroxyl, alkoxy, acyl,halogen, mercapto, oxo (═O), carboxyl, acyl, halo-substituted alkyl,amino, cyano, nitro, amidino, guanidino, an optionally substitutedcarbocycle or an optionally substituted heterocycle; wherein each alkylis independently a branched or unbranched, saturated or unsaturatedaliphatic hydrocarbon group, having up to 12 carbon atoms; or saltsthereof.
 2. The compound of claim 1, wherein Q is a carbocycle orheterocycle optionally substituted with one or more halogen, hydroxyl,cyano, amino, oxo, alkyl, a carbocycle or a heterocycle; wherein one ormore CH₂ groups of the alkyl substituent on Q is optionally replacedwith —O—, —S—, —S(O)—, —S(O)₂—, —N(R₈)—, —C(O)—, —C(O)—NR₈—, —NR₈—C(O)—,—SO₂—NR₈—, —NR₈—SO₂—, —NR₈—C(O)—NR₈—, —NR₈—C(NH)NR₈—, —NR₈—C(NH)—,—C(O)—O— or —O—C(O)—, and the alkyl is optionally substituted with oneor more halogen, cyano, or an optionally substituted carbocycle; andwherein said carbocycle or heterocycle substituents on Q are optionallysubstituted with one or more halogen, amino, hydroxyl, mercapto,carboxyl, or alkoxy.
 3. The compound of claim 1, wherein Q is selectedfrom the group consisting of IIIa-IIIs:

wherein n is 1-4; T is O, S, NR₈ or CR₇R₇; and W is O, NR₈ or CR₇R₇;wherein each R₇ is independently H, cyano, halogen, C₁₋₁₂alkyl, phenyl,or a monocyclic 5 to 14-membered aromatic or non-aromatic heterocyclehaving 1 to 4 heteroatoms selected from N, O, and S; wherein one or moreCH₂ groups of each alkyl are optionally replaced with —O—, —S(O)₂—,—C(O)—NR₈—, —NR₈—C(O)—, —C(O)—O— or —O—C(O)—; and each alkyl, phenyl, orheterocycle is optionally substituted with halogen; and wherein each R₈is independently H or methyl.
 4. The compound of claim 1, wherein R₁ isH.
 5. The compound of claim 1, wherein R₂ is alkyl, cycloalkyl or aheterocycle.
 6. The compound of claim 1, wherein R₂ is selected from thegroup consisting of t-butyl, isopropyl, cyclohexyl,tetrahydropyran-4-yl, N-methylsulfonylpiperidin-4-yl,tetrahydrothiopyran-4-yl, tetrahydrothiopyran-4-yl (in which the S is inoxidized form SO or SO₂), cyclohexan-4-one, 4-hydroxycyclohexane,4-hydroxy-4-methylcyclohexane, 1-methyl-tetrahydropyran-4-yl,2-hydroxyprop-2-yl, but-2-yl, phenyl and 1-hydroxyeth-1-yl.
 7. Thecompound of claim 1, wherein R₃ is methyl.
 8. The compound of claim 1,wherein R₄ is H or methyl.
 9. The compound of claim 1, wherein R₅ is Hor methyl.
 10. The compound of claim 1, wherein R₆ and R₆′ areindependently H or methyl.
 11. The compound of claim 1, wherein X₁ andX₂ are independently O.
 12. The compound of claim 1, wherein R₁ is H; R₂is isopropyl, t-butyl, cyclohexyl or tetrahydropyranyl; R₃ is methyl;R₃′ is H; R₄ is H or methyl; R₅ is H or methyl; and X₁ and X₂ are bothO.
 13. A compound selected from the group consisting of:

and salts thereof.